Akt protein kinase inhibitors

ABSTRACT

The present invention provides compounds, including resolved enantiomers, diastereomers, solvates and pharmaceutically acceptable salts thereof, comprising the Formula: 
       A-L-CR 
     where CR is a cyclical core group, L is a linking group and A is as defined herein. Also provided are methods of using the compounds of this invention as AKT protein kinase inhibitors and for the treatment of hyperproliferative diseases such as cancer.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No. 10/993,173filed on Nov. 19, 2004, which claims the benefit of U.S. ProvisionalApplication Ser. No. 60/524,003, filed Nov. 21, 2003, which areincorporated herein by reference in their entireties.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to novel inhibitors of serine/threonine proteinkinases (e.g., AKT and related kinases), pharmaceutical compositionscontaining the inhibitors, and methods for preparing these inhibitors.The inhibitors are useful for the treatment of hyperproliferativediseases, such as cancer and inflammation, in mammals and especially inhumans.

2. Description of the State of the Art

Protein kinases are a class of enzymes that catalyze the transfer of theγ-phosphorate group from ATP to a recipient protein, acting as asubstrate. The specific target of the kinase is the hydroxyl group of aserine, threonine or tyrosine residue. As a result of this specifictargeting, kinases are generally referred to as serine/threonine proteinkinases or tyrosine protein kinases. The human genome is estimated toencode in excess of 500 distinct protein kinases.

The seemingly insignificant task of phosphorylation of a serine,threonine or tyrosine residue belies the importance of protein kinasesin the processes of signal transduction and regulation of cellularfunctions. Kinases are typically mediated by transmembrane cellularreceptors, such as G-protein coupled receptors or growth factorreceptors, which when activated by extracellular ligands cause thephosphorylation of intracellular proteins. Often, an interconnectedseries (or cascade) of protein kinases is necessary to exert the overalleffect of this initial signal, which can ultimately result in effects asextreme as cell death (apoptosis).

The ratio of phosphorylated to unphosphorylated protein is a delicateequilibrium, with protein phosphatases acting as the negative regulatorof protein kinases, removing the phosphoryl group as it is no longerrequired. As an example of this interplay, the phosphorylation state ofkinases can control whether a cell undergoes division, arrests in thecell cycle or programmed cell death. Should this kinase/phosphataserelationship become disregulated, the potential consequences relating todisease are enormous. For example, abnormal protein kinase activity orexpression may be correlated with numerous hyperproliferative diseases,inflammation and tissue repair, and has been associated with a largenumber of diseases ranging from the relatively non-life threatening,such as psoriasis, to those which are almost always fatal, such asglioblastoma multiforme, an aggressive brain cancer.

Significantly, atypical protein phosphorylation and/or expression isoften reported to be one of the causative effects of abnormal cellularproliferation, metastasis and cell survival in cancer. The abnormalregulation and/or expression of various kinases, including VEGF, ILK,AKT, ROCK, p70S6K, Bcl, PKA, PKC, Raf, Src, PDK1, ErbB2, MEK, IKK, Cdk,EGFR, BAD, CHK1, CHK2 and GSK3 amongst numerous others, has beenspecifically implicated in cancer.

Recent data from the CDC indicate that cancer is the second most commoncause of death in the United States, with nearly a quarter of all deathsreported being attributable to malignant neoplasms (Anderson, NationalVital Statistics Report, 2001, 49 (11):1). Despite recent advances inthe understanding of the genesis, progression and treatment of cancer,much still needs to be done to improve the overall prognosis of cancerpatients.

The phosphatidylinositol 3′-OH kinase (PI3K) pathway is one of thesignaling pathways that exerts its effect on numerous cellular functionsincluding cell cycle progression, proliferation, motility, metabolismand survival. Activation of receptor protein tyrosine kinases (RTKs)cause PI3K to phosphorylate phosphatidylinositol (4,5)-diphosphate[PtdIns(4,5)P₂], generating the membrane-bound phosphatidylinositol(3,4,5)-triphosphate [PtdIns(3,4,5)P₃]. This in turn promotes therecruitment of a variety of protein kinases from the cytoplasm to theplasma membrane through the binding of PtdIns(3,4,5)P₃ to thepleckstrin-homology (PH) domain of the kinase. Kinases notable as keydownstream targets of PI3K include phosphoinositide-dependant kinase 1(PDK1) and AKT (also known as Protein Kinase B.) Phosphorylation of suchkinases then permits the activation or deactivation of numerous otherpathways involving mediators such as GSK3, mTOR, PRAS40, FKHD, NF-κB,BAD, Caspase-9, etc.

An important negative feedback mechanism for the PI3K pathway is PTEN, aphosphatase that catalyses the dephosphorylation of PtdIns(3,4,5)P₃ toPtdIns(4,5)P₂ (Furnari, F. B., et al, Cancer Res. 1998, 58:5002; Dahia,P. L. M., Hum. Molec. Genet. 1999, 8:185). It is of enormoussignificance that in greater than 60% of all solid tumors, PTEN ismutated into an inactive form, permitting the constitutive-activation ofthe PI3K pathway. As the majority of cancers are solid tumors, such anobservation would suggest that by specifically targeting either PI3Kitself or the individual downstream kinases in the PI3K pathway, onemight able to mitigate the effects of various cancers and restore normalcellular function.

One of the best-characterized targets of the PI3K lipid products is theAGC serine/threonine protein kinase AKT (Hemmings, B. A., Science, 1997,275:628). AKT is the human homologue of the protooncogene v-akt of theacutely transforming retrovirus AKT8. Its high sequence homology toprotein kinases A and C has also earned it the names Protein Kinase B(PKB) and Related to A and C(RAC.) Three isoforms of AKT are known toexist, namely Akt1, Akt2 and Akt3, which exhibit an overall homology of80% (Staal, S. P, Proc. Natl. Acad. Sci., 1987, 84:5034; Nakatani, K,Biochem. Biophys. Res. Commun., 1999, 257:906). In addition, both Akt2and Akt3 exhibit splice variants.

Upon recruitment to the cell membrane by PtdInd(3,4,5)P₃, AKT isphosphorylated (activated) by PDK1 at T308, T309 and T305 for isoformsAkt1, 2 and 3, respectively, and at S473, S474 and S472 for isoformsAkt1, 2 and 3, respectively. Such phosphorylation occurs by an as yetunknown kinase (putatively named PDK2), although PDK1 (Balendran, A.,Curr. Biol., 1999, 9:393), autophosphorylation (Toker, A., J. Biol.Chem., 2000, 275:8271) and integrin-linked kinase (ILK) (Delcommenne,M., Proc. Natl. Acad. Sci. USA, 1998, 95:11211) have been implicated inthis process. Although monophosphorylation of AKT activates the kinase,bis(phosphorylation) is required for maximal kinase activity.

AKT is believed to assert its effect on cancer by suppressing apoptosisand enhancing both angiogenesis and proliferation. In addition, AKT hasbeen shown to be overexpressed in many forms of human cancer including,but not limited to, colon (Zinda, et al, Clin. Cancer Res., 2001,7:2475), ovarian (Cheng, J. Q., et al., Proc. Natl. Acad. Sci. USA,1992, 89:9267), brain (Haas Kogan, D., et al, Curr. Biol., 1998,8:1195), lung (Brognard, J., et al, Cancer Res., 2001, 61:3986),pancreatic (Cheng, J. Q., et al., Proc. Natl. Acad. Sci., 1996,93:3636), prostate (Graff, J. R., et al, J. Biol. Chem., 2000,275:24500) and gastric carcinomas (Staal, S. P., et al., Proc. Natl.Acad. Sci. USA, 1987, 84:5034).

The development of kinase inhibitors that target abnormally regulatedpathways and ultimately result in disease is of enormous ethical andcommercial interest to the medical and pharmaceutical community. Assuch, a compound that inhibits (1) recruitment of AKT to the cellmembrane, (2) activation by PDK1 or PDK2, (3) substrate phosphorylation,or (4) one of the downstream targets of AKT would therefore be a validtarget as an anticancer agent, either as a stand-alone therapy or inconjunction with other accepted procedures.

SUMMARY OF THE INVENTION

This invention provides novel compounds that inhibit AKT proteinkinases, methods for producing these compounds, and pharmaceuticalcompositions containing such compounds. The compounds of the presentinvention have utility as therapeutic agents for diseases and conditionsthat can be treated by the inhibition of AKT protein kinases. Morespecifically, the present invention includes compounds, includingresolved enantiomers and diastereomers, and pharmaceutically acceptableprodrugs, metabolites, salts and solvates thereof, having the generalFormula I:

A-L-CR  (I)

where:

CR is heteroaryl, wherein said heteroaryl is optionally substituted withone or more groups selected from halogen, hydroxyl, cyano, nitro, azido,—NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹,—C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³,—NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴, —SO₂R²⁴, —NR²¹R²²,—NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³, —OR²¹, C₁-C₄ alkyl, C₁-C₆heteroalkyl, C₂-C₆ alkenyl, C₂-C₆ heteroalkenyl, C₂-C₆ alkynyl, C₂-C₆heteroalkynyl, C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, aryl,heteroaryl, arylalkyl and heteroarylalkyl, wherein any of said alkyl,heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,heterocycloalkyl, aryl, heteroaryl, arylalkyl and heteroarylalkyl arefurther optionally substituted with one or more groups selected fromhalogen, hydroxyl, cyano, nitro, azido, fluoromethyl, difluoromethyl,trifluoromethyl, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆heteroalkyl, C₂-C₆ heteroalkenyl, C₂-C₆ heteroalkynyl, C₃-C₆ cycloalkyl,C₃-C₆ heterocycloalkyl, —SR²¹, —S(O)R²⁴, —SO₂R²⁴, —C(O)R²¹, C(O)OR²¹,—C(O)NR²¹R²², —NR²¹R²² and —OR²¹;

L is selected from:

R¹² is hydrogen, halogen, hydroxy, cyano, nitro, amino, azido, C₁-C₅alkyl, C₂-C₅ alkenyl, C₂-C₅ alkynyl, C₁-C₅ heteroalkyl, C₂-C₅heteroalkenyl or C₂-C₅ heteroalkynyl, wherein any of said alkyl,alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl areoptionally substituted with one or more groups independently selectedfrom halogen, cyano, nitro, amino, azido, C₁-C₄ alkyl, fluoromethyl,difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy, difluoromethoxyand trifluoromethoxy;

A is

W is N or CR¹⁵, provided that when L is a substituted or unsubstitutedpiperazinylene, W must be CR¹⁵;

G is hydrogen, alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, whereinany of said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl isoptionally substituted with one or more groups selected from halogen,hydroxyl, cyano, amino, nitro, azido, —NR²¹SO₂R²⁴, —SO₂NR²¹R²²,—NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹, —C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹,—NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³, —NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹,—S(O)R²⁴, —SO₂R²⁴, —NR²¹R²², —NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³,—OR²¹, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, cycloalkyl,heterocycloalkyl aryl and heteroaryl;

B¹ and B² are independently absent or C₁-C₄ alkylene, C₁-C₄heteroalkylene, C₂-C₄ alkenylene, C₂-C₄ heteroalkenylene, C₂-C₄alkynylene, C₂-C₄ heteroalkynylene, C₃-C₆ cycloalkylene, and C₃-C₆heterocycloalkylene, wherein any of said alkylene, heteroalkylene,alkenylene, heteroalkenylene, alkynylene, heteroalkynylene,cycloalkylene or heterocycloalkylene is optionally substituted with oneor more groups independently selected from halogen, hydroxyl, cyano,nitro, azido, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, NR²¹R²² and OR²¹;

R²¹, R²² and R²³ independently are hydrogen, C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₁-C₆ heteroalkyl, C₂-C₆ heteroalkenyl, C₂-C₆heteroalkynyl, C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, aryl,arylalkyl, heteroaryl or heteroarylalkyl;

R²⁴ is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ heteroalkyl,C₂-C₆ heteroalkenyl, C₂-C₆ heteroalkynyl, C₃-C₆ cycloalkyl, C₃-C₆heterocycloalkyl, aryl, arylalkyl, heteroaryl or heteroarylalkyl;

or any two of R²¹, R²², R²³ or R²⁴ together with the atom(s) to whichthey are attached form a 4 to 10 membered carbocyclic, aryl, heteroarylor heterocyclic ring, wherein any of said carbocyclic, aryl, heteroarylor heterocyclic rings are optionally substituted with one or more groupsindependently selected from halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl;

R¹³ and R¹⁴ are independently hydrogen, hydroxyl, cyano, C₁-C₆ alkyl,C₁-C₆ heteroalkyl, C₂-C₆ alkenyl, C₂-C₆ heteroalkenyl, C₂-C₆ alkynyl,C₂-C₆ heteroalkynyl, C₁-C₆ cycloalkyl, C₁-C₆ heterocycloalkyl, aryl,heteroaryl, arylalkyl, heteroarylalkyl, —C(O)R²¹, C(O)OR²¹,C(═NR²¹)NR²²R²³ or —SO₂R²⁴, wherein any of said alkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, aryl, heteroaryl, heteroalkyl,heteroalkenyl, heteroalkynyl, arylalkyl or heteroarylalkyl is optionallysubstituted with one or more groups independently selected from halogen,hydroxyl, cyano, amino, nitro, azido, —NR²¹SO₂R²⁴, —SO₂NR²¹R²²,—NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹, —C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹,—NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³, —NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹,—S(O)R²⁴, —SO₂R²⁴, —NR²¹R²², —NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³,—OR²¹, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, cycloalkyl,heterocycloalkyl aryl and heteroaryl;

or R¹³ and R¹⁴ together with the atoms to which they are attached form a4 to 10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring,wherein any of said carbocyclic, aryl, heteroaryl and heterocyclic ringsare optionally substituted with one or more groups independentlyselected from halogen, cyano, nitro, azido, C₁-C₄ alkyl, C₁-C₄heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄heteroalkynyl, NR²¹R²² and OR²¹;

or R¹³ and an atom of B² together with N form a 4 to 10 memberedcarbocyclic, aryl, heteroaryl or heterocyclic ring, wherein any of saidcarbocyclic, aryl, heteroaryl and heterocyclic rings are optionallysubstituted with one or more groups independently selected from halogen,cyano, nitro, azido, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl,C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, NR²¹R²² andOR²¹;

R¹⁵ is hydrogen, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl or C₂-C₄ heteroalkynyl, wherein any of saidalkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl or heteroalkynyl isoptionally substituted with one or more groups independently selectedfrom halogen, hydroxyl, cyano, nitro, azido, NR²¹R²² and OR²¹;

or R¹³ and R¹⁵ together with atoms to which they are attached form a 3to 10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring,wherein any of said carbocyclic, aryl, heteroaryl and heterocyclic ringsare optionally substituted with one or more groups independentlyselected from halogen, cyano, nitro, azido, C₁-C₄ alkyl, C₁-C₄heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄heteroalkynyl, NR²¹R²² and OR²¹;

or, when W is CR¹⁵, R¹⁵ and an atom of B¹ or B² together with C, form a3 to 10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring,wherein any of said carbocyclic, aryl, heteroaryl and heterocyclic ringsare optionally substituted with one or more groups independentlyselected from halogen, cyano, nitro, azido, C₁-C₄ alkyl, C₁-C₄heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄heteroalkynyl, NR²¹R²² and OR²¹.

The invention also relates to pharmaceutical compositions comprising aneffective amount of an agent selected from compounds of Formula I.Methods of making the compounds of Formula I are also described.

In a further embodiment, the present invention provides methods ofinhibiting the activity of AKT protein kinases utilizing compounds ofFormula I.

In a further embodiment, the present invention provides a method oftreating diseases or medical conditions mediated by AKT protein kinases.For example, this invention provides a method for treatment of ahyperproliferative disorder in a warm-blooded animal which comprisesadministering to such animal one or more compounds of Formula I, or apharmaceutically acceptable salt or in vivo cleavable prodrug thereof inan amount effective to treat or prevent said hyperproliferativedisorder.

In a further embodiment, the present invention provides a method ofinhibiting the production of AKT protein kinases, which comprisesadministering to a warm-blooded animal an effective amount of a compoundof Formula I, or a pharmaceutically acceptable salt or in vivo cleavableprodrug thereof in an amount effective to inhibit production of an AKTprotein kinase.

In a further embodiment, the present invention provides a method ofproviding AKT protein kinase inhibiting effect comprising administeringto a warm-blooded animal an effective amount of a compound of Formula I,or a pharmaceutically acceptable salt or in vivo cleavable prodrugthereof.

In a further embodiment, the present invention provides treating orpreventing an AKT protein kinase mediated condition, comprisingadministering to a mammal a compound having Formula I or apharmaceutically-acceptable salt, in vivo cleavable prodrug orpharmaceutical formulation thereof, in an amount effective to treat orprevent said AKT protein kinase-mediated condition. AKT protein kinasemediated conditions that can be treated according to the methods of thisinvention include, but are not limited to, cancer, inflammation andvarious proliferative, cardiovascular, neurodegenerative, gynecological& dermatological diseases.

Hyperproliferative conditions that can be treated according to themethods of this invention include, but are not limited to, cancers ofthe head, neck, lung, breast, colon, ovary, bladder, stomach, esophagus,uterus or prostate, among other kinds of hyperproliferative disorders.In compounds and methods of this invention can be used to treat diseasesand conditions, including rheumatoid arthritis, osteoarthritis,endometriosis, atherosclerosis, vein graft stenosis, peri-anastomaticprothetic graft stenosis, prostate hyperplasia, chronic obstructivepulmonary disease, psoriasis, inhibition of neurological damage due totissue repair, scar tissue formation (and can aid in wound healing),multiple sclerosis, inflammatory bowel disease, infections, particularlybacterial, viral, retroviral or parasitic infections (by increasingapoptosis), pulmonary disease, neoplasm, Parkinson's disease, transplantrejection (as an immunosupressant), macular degeneration and septicshock.

The compounds of Formula I may be used advantageously in combinationwith other known therapeutic agents.

Additional advantages and novel features of this invention shall be setforth in part in the description that follows, and in part will becomeapparent to those skilled in the art upon examination of the followingspecification, or may be learned by the practice of the invention. Theadvantages of the invention may be realized and attained by means of theinstrumentalities, combinations, compositions, and methods particularlypointed out in the appended claims.

BRIEF DESCRIPTION OF THE FIGURES

The accompanying drawings, which are incorporated herein and form a partof the specification, illustrate non-limiting embodiments of the presentinvention, and together with the description, serve to explain theprinciples of the invention.

In the Figures:

FIG. 1 shows a reaction scheme for the preparation of compounds 8-11.

FIG. 2 shows a reaction scheme for the preparation of compound 14.

FIG. 3 shows a reaction scheme for the preparation of compound 18.

FIG. 4 shows a reaction scheme for the preparation of compound 22.

FIG. 5 shows a reaction scheme for the preparation of compound 28.

FIG. 6 shows a reaction scheme for the preparation of compound 31.

FIG. 7 shows a reaction scheme for the preparation of compounds 35 and36.

FIG. 8 shows a reaction scheme for the preparation of compounds 39 and40.

FIG. 9 shows a reaction scheme for the preparation of compound 40.

FIG. 10 shows a reaction scheme for the preparation of compound 46.

FIG. 11 shows a reaction scheme for the preparation of compound 50.

FIG. 12 shows a reaction scheme for the preparation of compound 55.

FIG. 13 shows a reaction scheme for the preparation of compounds 57 and58.

FIG. 14 shows a reaction scheme for the preparation of compounds 60 and61.

FIG. 15 shows a reaction scheme for the preparation of compounds 69-71.

FIG. 16 shows a reaction scheme for the preparation of compounds 72-74.

FIG. 17 shows a reaction scheme for the preparation of compound 78.

FIG. 18 shows a reaction scheme for the preparation of compounds 80 and81.

FIG. 19 shows a reaction scheme for the preparation of compounds 85 and86.

FIG. 20 shows a reaction scheme for the preparation of compound 90.

FIG. 21 shows a reaction scheme for the preparation of compounds 93-97.

FIG. 22 shows a reaction scheme for the preparation of compounds 100 and101.

FIG. 23 shows a reaction scheme for the preparation of compounds104-109.

FIG. 24 shows a reaction scheme for the preparation of compounds112-116.

FIG. 25 shows a reaction scheme for the preparation of compounds120-125.

FIG. 26 shows a reaction scheme for the preparation of compounds 127 and129.

FIG. 27 shows a reaction scheme for the preparation of compounds 132 and134.

FIG. 28 shows a reaction scheme for the preparation of compounds 137 and139.

FIG. 29 shows a reaction scheme for the preparation of compounds141-144.

FIG. 30 shows a reaction scheme for the preparation of compound 148.

FIG. 31 shows a reaction scheme for the preparation of compounds151-153.

FIG. 32 shows a reaction scheme for the preparation of compounds 155 and156.

FIG. 33 shows a reaction scheme for the preparation of compounds 161 and162.

FIG. 34 shows a reaction scheme for the preparation of compounds171-175.

FIG. 35 shows a reaction scheme for the preparation of compounds178-182.

FIG. 36 shows a reaction scheme for the preparation of compounds 179.

FIG. 37 shows a reaction scheme for the preparation of compound 190.

FIG. 38 shows a reaction scheme for the preparation of compounds197-199.

FIG. 39 shows a reaction scheme for the preparation of compounds205-208.

FIG. 40 shows a reaction scheme for the preparation of compounds 215 and217.

FIG. 41 shows a reaction scheme for the preparation of compounds 219,221 and 223.

FIG. 42 shows a reaction scheme for the preparation of compound 229.

FIG. 43 shows a reaction scheme for the preparation of compounds 232 and234.

FIG. 44 shows a reaction scheme for the preparation of compounds237-242.

FIG. 45 shows a reaction scheme for the preparation of compounds 244 and247.

FIG. 46 shows a reaction scheme for the preparation of compounds 250,251 and 254-256.

FIG. 47 shows a reaction scheme for the preparation of compounds 263 and265.

FIG. 48 shows a reaction scheme for the preparation of compounds 269 and271.

DETAILED DESCRIPTION OF THE INVENTION

The inventive compounds of Formula I are useful for inhibiting AKTprotein kinases. The compounds of Formula I may also be useful asinhibitors of tyrosine kinases as well as serine and threonine kinasesin addition to AKT. Such compounds have utility as therapeutic agentsfor diseases that can be treated by the inhibition of the AKT proteinkinase signaling pathway and tyrosine and serine/threonine kinasereceptor pathways. In general, the invention includes compounds,including resolved enantiomers and diastereomers, and pharmaceuticallyacceptable prodrugs, metabolites, salts and solvates thereof, having thegeneral Formula I:

AA-L-CR  I

where CR is heteroaryl, wherein said heteroaryl is optionallysubstituted with one or more groups selected from halogen, hydroxyl,cyano, nitro, azido, —NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴,—S(O)NR²¹R²², —C(O)R²¹, —C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, —NR²¹C(O)OR²⁴,—NR²¹C(═NR²¹)NR²²R²³, —NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴,—SO₂R²⁴, —NR²¹R²², —NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³, —OR²¹, C₁-C₄alkyl, C₁-C₆ heteroalkyl, C₂-C₆ alkenyl, C₂-C₆ heteroalkenyl, C₂-C₆alkynyl, C₂-C₆ heteroalkynyl, C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl,aryl, heteroaryl, arylalkyl and heteroarylalkyl, wherein any of saidalkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl,cycloalkyl, heterocycloalkyl, aryl, heteroaryl, arylalkyl andheteroarylalkyl are further optionally substituted with one or moregroups selected from halogen, hydroxyl, cyano, nitro, azido,fluoromethyl, difluoromethyl, trifluoromethyl, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₁-C₆ heteroalkyl, C₂-C₆ heteroalkenyl, C₂-C₆heteroalkynyl, C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, —SR²¹,—S(O)R²⁴, —SO₂R²⁴, —C(O)R²¹, C(O)OR²¹, —C(O)NR²¹R²², —NR²¹R²² and —OR²¹;

L is selected from:

R¹² is hydrogen, halogen, hydroxy, cyano, nitro, amino, azido, C₁-C₅alkyl, C₂-C₅ alkenyl, C₂-C₅ alkynyl, C₁-C₅ heteroalkyl, C₂-C₅heteroalkenyl or C₂-C₅ heteroalkynyl, wherein any of said alkyl,alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl areoptionally substituted with one or more groups independently selectedfrom halogen, cyano, nitro, amino, azido, C₁-C₄ alkyl, fluoromethyl,difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy, difluoromethoxyand trifluoromethoxy;

A is

W is N or CR¹⁵, provided that when L is a substituted or unsubstitutedpiperazinylene, W must be CR¹⁵;

G is hydrogen, alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, whereinany of said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl isoptionally substituted with one or more groups selected from halogen,hydroxyl, cyano, amino, nitro, azido, —NR²¹SO₂R²⁴, —SO₂NR²¹R²²,—NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹, —C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹,—NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³, —NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹,—S(O)R²⁴, —SO₂R²⁴, —N²¹R²², —NR²¹C(O)NR²²R²³, —NR²C(NCN)NR²²R²³, —OR²¹,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl,C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, cycloalkyl, heterocycloalkyl aryland heteroaryl;

B¹ and B² are independently absent or C₁-C₄ alkylene, C₁-C₄heteroalkylene, C₂-C₄ alkenylene, C₂-C₄ heteroalkenylene, C₂-C₄alkynylene, C₂-C₄ heteroalkynylene, C₃-C₆ cycloalkylene, and C₃-C₆heterocycloalkylene, wherein any of said alkylene, heteroalkylene,alkenylene, heteroalkenylene, alkynylene, heteroalkynylene,cycloalkylene or heterocycloalkylene is optionally substituted with oneor more groups independently selected from halogen, hydroxyl, cyano,nitro, azido, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, NR²¹R²² and OR²¹;

R²¹, R²² and R²³ independently are hydrogen, C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₁-C₆ heteroalkyl, C₂-C₆ heteroalkenyl, C₂-C₆heteroalkynyl, C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, aryl,arylalkyl, heteroaryl or heteroarylalkyl;

R²⁴ is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ heteroalkyl,C₂-C₆ heteroalkenyl, C₂-C₆ heteroalkynyl, C₃-C₆ cycloalkyl, C₃-C₆heterocycloalkyl, aryl, arylalkyl, heteroaryl or heteroarylalkyl;

or any two of R²¹, R²², R²³ or R²⁴ together with the atom(s) to whichthey are attached form a 4 to 10 membered carbocyclic, aryl, heteroarylor heterocyclic ring, wherein any of said carbocyclic, aryl, heteroarylor heterocyclic rings are optionally substituted with one or more groupsindependently selected from halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl;

R¹³ and R¹⁴ are independently hydrogen, hydroxyl, cyano, C₁-C₆ alkyl,C₁-C₆ heteroalkyl, C₂-C₆ alkenyl, C₂-C₆ heteroalkenyl, C₂-C₆ alkynyl,C₂-C₆ heteroalkynyl, C₁-C₆ cycloalkyl, C₁-C₆ heterocycloalkyl, aryl,heteroaryl, arylalkyl, heteroarylalkyl, —C(O)R²¹, C(O)OR²¹,C(═NR²¹)NR²²R²³ or —SO₂R²⁴, wherein any of said alkyl, alkenyl, alkynyl,cycloalkyl, heterocycloalkyl, aryl, heteroaryl, heteroalkyl,heteroalkenyl, heteroalkynyl, arylalkyl or heteroarylalkyl is optionallysubstituted with one or more groups independently selected from halogen,hydroxyl, cyano, amino, nitro, azido, —NR²¹SO₂R²⁴, —SO₂NR²¹R²²,—NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹, —C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹,NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³, —NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹,—S(O)R²⁴, —SO₂R²⁴, —NR²¹R²², —NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³,—OR²¹, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, cycloalkyl,heterocycloalkyl aryl and heteroaryl;

or R¹³ and R¹⁴ together with the atoms to which they are attached form a4 to 10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring,wherein any of said carbocyclic, aryl, heteroaryl and heterocyclic ringsare optionally substituted with one or more groups independentlyselected from halogen, cyano, nitro, azido, C₁-C₄ alkyl, C₁-C₄heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄heteroalkynyl, NR²¹R²² and OR²¹;

or R¹³ and an atom of B² together with N form a 4 to 10 memberedcarbocyclic, aryl, heteroaryl or heterocyclic ring, wherein any of saidcarbocyclic, aryl, heteroaryl and heterocyclic rings are optionallysubstituted with one or more groups independently selected from halogen,cyano, nitro, azido, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl,C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, NR²¹R²² andOR²¹;

R¹⁵ is hydrogen, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl or C₂-C₄ heteroalkynyl, wherein any of saidalkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl or heteroalkynyl isoptionally substituted with one or more groups independently selectedfrom halogen, hydroxyl, cyano, nitro, azido, NR²¹R²² and OR²¹;

or R¹³ and R¹⁵ together with atoms to which they are attached form a 3to 10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring,wherein any of said carbocyclic, aryl, heteroaryl and heterocyclic ringsare optionally substituted with one or more groups independentlyselected from halogen, cyano, nitro, azido, C₁-C₄ alkyl, C₁-C₄heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄heteroalkynyl, NR²¹R²² and OR²¹;

or, when W is CR¹⁵, R¹⁵ and an atom of B¹ or B² together with C, form a3 to 10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring,wherein any of said carbocyclic, aryl, heteroaryl and heterocyclic ringsare optionally substituted with one or more groups independentlyselected from halogen, cyano, nitro, azido, C₁-C₄ alkyl, C₁-C₄heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄heteroalkynyl, NR²¹R²² and OR²¹.

In one embodiment of the invention, CR is selected from:

where X is N or CR¹;

Y is CR² or N, provided that when X is N, Y must be CR²;

Z is CR³R^(3a) or NR^(2a), provided that when X is N, Z must be CR³;

D¹, D², D³ and D⁴ are independently CR⁴ or N, provided that no more thantwo of D¹, D², D³ or D⁴ are N;

- - - - - is an optional double bond;

D⁵ is CR⁵R^(5a), NR^(2a), O or S, provided that when D⁵ is O or S, D⁸must be C, D⁷ must be CR⁷ or N, and either (i) Q must be CR⁶ orCR⁶R^(6a) or (ii) D⁷ must be CR⁷ or CR⁷R^(7a);

Q is CR⁶, N or C═O, provided that either (w) when Q is N, one of D⁵, D⁷and D⁸ must be C, or (x) when Q is C═O, D⁵ must be CR⁵ or N, D⁷ must beCR⁷ or N, and D⁸ must be C;

D⁷ is CR⁷, N, O or S, provided that when D⁷ is O or S, D⁸ must be C, D⁵must be CR⁵ or N, and either (y) Q must be CR⁶, or (z) D⁵ must be CR⁵;

D⁸ is C or N, provided that when D⁸ is N, D⁵ must be CR⁵R^(5a) and Qmust be CR⁶ or CR⁶R^(6a);

either K or M is carbonyl, provided that both K and M are not carbonyl;

R¹, R⁵, R^(5a) and R⁸ are independently hydrogen, halogen, hydroxyl,cyano, nitro, azido, fluoromethyl, difluoromethyl, trifluoromethyl,methoxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy, amino,aminomethyl, dimethylamino, aminoethyl, diethylamino or ethoxy;

R² is hydrogen, halogen, hydroxyl, cyano, nitro, amino, azido, C₁-C₄alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄alkynyl, C₂-C₄ heteroalkynyl, C₁-C₆ cycloalkyl, C₁-C₆ heterocycloalkyl,C₁-C₆ aryl, or C₁-C₆ heteroaryl, wherein any of said alkyl, heteroalkyl,alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,heterocycloalkyl, aryl or heteroaryl are further optionally substitutedwith one or more groups independently selected from halogen, hydroxyl,cyano, nitro, azido, fluoromethyl, difluoromethyl, trifluoromethyl,methoxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy, amino,aminomethyl, dimethylamino, aminoethyl, diethylamino and ethoxy;

R^(2a) is hydrogen, hydroxyl, cyano, C₁-C₄ alkyl, C₁-C₄ heteroalkyl,C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₆ heteroalkynyl,C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, aryl or heteroaryl, whereinany of said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl areoptionally substituted with one or more groups independently selectedfrom halogen, hydroxyl, cyano, nitro, azido fluoromethyl,difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy,difluoromethoxy, trifluoromethoxy, amino, aminomethyl, dimethylamino,aminoethyl, diethylamino or ethoxy;

R³ and R^(3a) are independently hydrogen, halogen, hydroxyl, cyano,nitro, amino azido, C₁-C₆ alkyl, C₁-C₆ heteroalkyl, C₂-C₆ alkenyl, C₂-C₆heteroalkenyl, C₂-C₆ alkynyl, C₂-C₆ heteroalkynyl, C₁-C₆ cycloalkyl,C₁-C₆ heterocycloalkyl, aryl or heteroaryl, wherein any of said alkyl,heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,heterocycloalkyl, aryl or heteroaryl are further optionally substitutedwith one or more groups independently selected from halogen, hydroxyl,cyano, nitro, azido, OR¹, NR¹R², and (C═O)R²;

R⁴, R⁶, R^(6a), R⁷, R^(7a) and R¹⁰ are independently hydrogen, hydroxyl,cyano, amino, nitro, azido, alkyl, heteroalkyl, alkenyl, heteroalkenyl,alkynyl, heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, —NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴,—S(O)NR²¹R²², —C(O)R²¹, —C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, —NR²¹C(O)OR²⁴,—NR²¹C(NR²¹)NR²²R²³, —NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴,—SO₂R²⁴, —NR²¹R²², NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³ or —OR²¹, whereinany of said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroarylor heteroarylalkyl, is optionally substituted with one or more groupsindependently selected from halogen, hydroxyl, cyano, amino, nitro,azido, —NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹,—C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, —NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³,—NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴, —SO₂R²⁴, —NR²¹R²²,—NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³, —OR²¹, C₁-C₄ alkyl, C₁-C₄heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, andwherein said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl mayoptionally be further optionally substituted with one or more groupsindependently selected from halogen, hydroxyl, cyano, amino, nitro,azido, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, cycloalkyl,heterocycloalkyl, aryl and heteroaryl, —NR²¹R²² and —OR²¹;

or R⁶ and R⁷ together with the atoms to which they are attached form a 4to 10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring,wherein any of said carbocyclic, aryl, heteroaryl and heterocyclic ringsare optionally substituted with one or more groups independentlyselected from halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, OR¹, NR¹R², cycloalkyl, heterocycloalkyl, aryl,heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl; and

R⁹ is hydrogen, halogen, hydroxyl, cyano, nitro, azido, C₁-C₆ alkyl,C₁-C₆ heteroalkyl, C₂-C₆ alkenyl, C₂-C₆ heteroalkenyl, C₂-C₆ alkynyl,C₂-C₆ heteroalkynyl, C₁-C₆ cycloalkyl, C₁-C₆ heterocycloalkyl, aryl,heteroaryl, —NR²¹R²², —OR²¹, —NR²¹SO₂R²⁴ and —NR²¹C(O)R²², wherein anyof said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl arefurther optionally substituted with one or more groups independentlyselected from halogen, hydroxyl, cyano, nitro, azido, OR¹, NR¹R², and(C═O)R².

In one embodiment of the invention, CR is selected from

where D⁵, D⁷, D⁸, X and Q are as defined above.

According to another embodiment of the invention, A is:

where B¹ and B² are, independently, absent or C₁-C₄ alkylene;

R^(21a)—R^(21c) are independently H, halogen, CH₃, CF₃, CH₃O, CN, NO₂,NH₂, Ph, OH, or OCH₂Ph;

R^(22a), R^(22b), and R²⁴ are independently H, CH₃, or halogen;

R^(23a) is H; and

R^(23b) is H, CH₃, CH₂NH₂, CH₂NHCH₂, CH₂CH₂NH₂, CH₂CH₂NHCH₂,CH₂CH₂N(CH₂)₂, —(C═O)CH₂NH₂ or —(C═O)CH₂CH₂NH₂;

or R^(23a) and R^(23b) are joined to complete a 5 or 6 memberedheterocyclic ring.

In yet another embodiment of this invention, A is

where R²⁵ and R²⁶ are independently H or CH₃; and

R²⁷ is 1-naphthyl, 2-naphthyl, 3′-benzylthienyl, 2′-thienyl, 2′-pyridyl,3′-pyridyl, 4′-pyridyl, 4′-thiazolyl, or 3,3-diphenyl.

More specific examples of the A group of Formula I according to thisinvention include, but are not limited to,

where G, B¹, R¹³ and R¹⁴ are as defined above.

In an alternative embodiment, the A group of Formula I of this inventionis a D- or L-amino acid selected from the 20 naturally occurring aminoacids commonly designated by three letter symbols, and also includesunnatural amino acids including, but not limited to, 4-hydroxyproline,hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvaline,beta-alanine, gamma-aminobutyric acid, cirtulline, homocysteine,homoserine, ornithine and methionine sulfone. In one preferredembodiment, the A group of Formula I is alanine, phenylalanine,histidine, or tryptophan.

A specific example of a compound based on Formula I is:

Another example of a compound based on Formula I is:

wherein R²⁸ is H, halogen, CH₃, CF₃, CH₃O, CN, NO₂, NH₂, Ph, OH, orOCH₂Ph.

Still another example of a compound based on Formula I is:

where D¹⁶ is O or N; and

R²⁹ is H, halogen, CH₃, CF₃, CH₃O, CN, NO₂, NH₂, Ph, OH, or OCH₂Ph.

The term “alkyl” as used herein refers to a saturated linear orbranched-chain monovalent hydrocarbon radical of one to twelve carbonatoms, wherein the alkyl radical may be optionally substitutedindependently with one or more substituents described below. Examples ofalkyl groups include, but are not limited to, methyl, ethyl, n-propyl,isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl,tert-pentyl, hexyl, isohexyl, and the like.

The term “alkylene” as used herein refers to a linear or branched-chainsaturated divalent hydrocarbon radical of one to twelve carbon atoms,e.g., methylene, ethylene, propylene, 2-methylpropylene, pentylene, andthe like. The alkylene radical may be optionally substitutedindependently with one or more substituents described herein.

The term “heteroalkyl” refers to saturated linear or branched-chainmonovalent hydrocarbon radical of one to twelve carbon atoms, wherein atleast one of the carbon atoms is replaced with a heteroatom selectedfrom N, O, or S, and wherein the radical may be a carbon radical orheteroatom radical (i.e., the heteroatom may appear in the middle or atthe end of the radical). The heteroalkyl radical may be optionallysubstituted independently with one or more substituents describedherein. The term “heteroalkyl” encompasses alkoxy and heteroalkoxyradicals.

The term “heteroalkylene” as used herein refers to a linear orbranched-chain saturated divalent hydrocarbon radical of two to twelvecarbon atoms, wherein at least one of the carbon atoms is replaced witha heteroatom selected from N, O, or S, and wherein the radical may be acarbon radical or heteroatom radical (i.e., the heteroatom may appear inthe middle or at the end of the radical). The heteroalkylene radical maybe optionally substituted independently with one or more substituentsdescribed herein.

“Alkenyl” means a linear or branched-chain monovalent hydrocarbonradical of two to twelve carbon atoms containing at least one doublebond, wherein the alkenyl radical may be optionally substitutedindependently with one or more substituents described below. Examples ofalkenyl groups include, but are not limited to: ethylene or vinyl(—CH═CH₂), allyl (—CH₂CH═CH₂), 1-cyclopent-1-enyl, 1-cyclopent-2-enyl,1-cyclopent-3-enyl, 5-hexenyl (—CH₂ CH₂CH₂CH₂CH═CH₂), 1-cyclohex-1-enyl,1-cyclohex-2-enyl, and 1-cyclohex-3-enyl.

“Alkenylene” refers to an a linear or branched-chain divalenthydrocarbon radical of one to twelve carbon atoms containing at leastone double bond, e.g., 1,2-ethylene (—CH═CH—). The alkenylene radicalmay be optionally substituted independently with one or moresubstituents described herein.

The term “heteroalkenyl” refers to a linear or branched-chain monovalenthydrocarbon radical of two to twelve carbon atoms and at least onedouble bond, wherein at least one of the carbon atoms is replaced with aheteroatom selected from N, O, or S, and wherein the radical may be acarbon radical or heteroatom radical (i.e., the heteroatom may appear inthe middle or at the end of the radical). The heteroalkenyl radical maybe optionally substituted independently with one or more substituentsdescribed herein. The term “heteroalkenyl” encompasses alkenoxy andheteroalkenoxy radicals.

“Heteroalkenylene” refers to an a linear or branched saturated divalenthydrocarbon radical of one to twelve carbon atoms containing at leastone double bond, wherein at least one of the carbon atoms is replacedwith a heteroatom selected from N, O, or S, and wherein the radical maybe a carbon radical or heteroatom radical (i.e., the heteroatom mayappear in the middle or at the end of the radical). The heteroalkenyleneradical may be optionally substituted independently with one or moresubstituents described herein.

The term “allyl” refers to a radical having the formula RC═CHCHR,wherein R is alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl,aryl, heteroaryl, or any substituent as defined herein, wherein theallyl radical may be optionally substituted independently with one ormore substituents described herein.

The term “alkynyl” means a linear or branched-chain monovalenthydrocarbon radical of two to twelve carbon atoms containing at leastone triple bond, wherein the alkynyl radical may be optionallysubstituted independently with one or more substituents described below.Examples of alkynyl groups include, but are not limited to: acetylene(—C≡CH) and propargyl (—CH₂C≡CH).

“Alkynylene” refers to a linear or branched-chain divalent hydrocarbonradical of one to twelve carbon atoms containing at least one triplebond. The alkynylene radical may be optionally substituted independentlywith one or more substituents described herein. Typical alkynyleneradicals include, but are not limited to: acetylene (—C≡C—), propargyl(—CH₂C≡C—), and 4-pentynyl (—CH₂CH₂CH₂C≡CH—).

The term “heteroalkynyl” refers to a linear or branched-chain monovalenthydrocarbon radical of two to twelve carbon atoms containing at leastone triple bond, wherein at least one of the carbon atoms is replacedwith a heteroatom selected from N, O, or S, and wherein the radical maybe a carbon radical or heteroatom radical (i.e., the heteroatom mayappear in the middle or at the end of the radical). The heteroalkynylradical may be optionally substituted independently with one or moresubstituents described herein. The term “heteroalkynyl” encompassesalkynoxy and heteroalkynoxy radicals.

The term “heteroalkynylene” refers to a linear or branched divalenthydrocarbon radical of two to twelve carbons containing at least onetriple bond, wherein at least one of the carbon atoms is replaced with aheteroatom selected from N, O, or S, and wherein the radical may be acarbon radical or heteroatom radical (i.e., the heteroatom may appear inthe middle or at the end of the radical). The heteroalkynylene radicalmay be optionally substituted independently with one or moresubstituents described herein.

The terms “carbocycle,” “carbocyclyl,” or “cycloalkyl” refer tosaturated or partially unsaturated cyclic hydrocarbon radical havingfrom three to ten carbon atoms. The term “cycloalkyl” includesmonocyclic and polycyclic (e.g., bicyclic and tricyclic) cycloalkylstructures, wherein the polycyclic structures optionally include asaturated or partially unsaturated cycloalkyl fused to a saturated orpartially unsaturated cycloalkyl or heterocycloalkyl ring or an aryl orheteroaryl ring. Examples of cycloalkyl groups include, but are notlimited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, and the like. The cycloalkyl may be optionally substitutedindependently in one or more substitutable positions with variousgroups. For example, such cycloalkyl groups may be optionallysubstituted with, for example, C₁-C₆ alkyl, C₁-C₆ alkoxy, halogen,hydroxy, cyano, nitro, amino, mono(C₁-C₆)alkylamino,di(C₁-C₆)alkylamino, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆ haloalkyl, C₁-C₆haloalkoxy, amino(C₁-C₆)alkyl, mono(C₁-C₆)alkylamino(C₁-C₆)alkyl ordi(C₁-C₆)alkylamino(C₁-C₆)alkyl.

The terms “heterocycloalkyl,” “heterocycle” or “heterocyclyl” refer to asaturated or partially unsaturated carbocyclic radical of 3 to 8 ringatoms in which at least one ring atom is a heteroatom selected fromnitrogen, oxygen and sulfur, the remaining ring atoms being C, where oneor more ring atoms may be optionally substituted independently with oneor more substituent described below. The radical may be a carbon radicalor heteroatom radical. The term further includes bicyclic and tricyclicfused ring systems which include a heterocycle fused one or morecarbocyclic or heterocyclic rings. “Heterocycloalkyl” also includesradicals where heterocycle radicals are fused with aromatic orheteroaromatic rings. Examples of heterocycloalkyl rings include, butare not limited to, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl,tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl,tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino,thioxanyl, piperazinyl, homopiperazinyl, azetidinyl, oxetanyl,thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl,thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl,indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl,pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl,dihydrofuranyl, pyrazolidinylimidazolinyl, imidazolidinyl,3-azabicyco[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl,azabicyclo[2.2.2]hexanyl, 3H-indolyl and quinolizinyl. Spiro moietiesare also included within the scope of this definition. The foregoinggroups, as derived from the groups listed above, may be C-attached orN-attached where such is possible. For instance, a group derived frompyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached).Further, a group derived from imidazole may be imidazol-1-yl(N-attached) or imidazol-3-yl (C-attached). An example of a heterocyclicgroup wherein 2 ring carbon atoms are substituted with oxo (═O) moietiesis 1,1-dioxo-thiomorpholinyl. The heterocycle groups herein areunsubstituted or, as specified, substituted in one or more substitutablepositions with various groups. For example, such heterocycle groups maybe optionally substituted with, for example, C₁-C₆ alkyl, C₁-C₆ alkoxy,halogen, hydroxy, cyano, nitro, amino, mono(C₁-C₆)alkylamino,di(C₁-C₆)alkylamino, C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₆ haloalkyl, C₁-C₆haloalkoxy, amino(C₁-C₆)alkyl, mono(C₁-C₆)alkylamino(C₁-C₆)alkyl ordi(C₁-C₆)alkylamino(C₁-C₆)alkyl.

The terms “heterocycloalkylene” refers to a saturated or partiallyunsaturated divalent carbocyclic radical of 3 to 8 ring atoms in whichat least one ring atom is a heteroatom selected from nitrogen, oxygenand sulfur, the remaining ring atoms being C, where one or more ringatoms may be optionally substituted independently with one or moresubstituent described herein. Examples include, but are not limited to,substituted and unsubstituted piperidinylenes.

The term “aryl” refers to a monovalent aromatic carbocyclic radicalhaving a single ring (e.g., phenyl), multiple rings (e.g., biphenyl), ormultiple condensed rings in which at least one is aromatic, (e.g.,1,2,3,4-tetrahydronaphthyl, naphthyl), which is optionally mono-, di-,or trisubstituted with, e.g., halogen, lower alkyl, lower alkoxy,trifluoromethyl, aryl, heteroaryl, and hydroxy.

The term “heteroaryl” refers to a monovalent aromatic radical of 5-, 6-,or 7-membered rings which includes fused ring systems (at least one ofwhich is aromatic) of 5-10 atoms containing at least one and up to fourheteroatoms selected from nitrogen, oxygen, or sulfur. Examples ofheteroaryl groups are pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl,triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl,oxazolyl, isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl, indolyl,benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indolizinyl,phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl, purinyl,oxadiazolyl, triazolyl, thiadiazolyl, thiadiazolyl, furazanyl,benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoxazolyl,quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl. Spiromoieties are also included within the scope of this definition.Heteroaryl groups are optionally mono-, di-, or trisubstituted with,e.g., halogen, lower alkyl, lower alkoxy, haloalkyl, aryl, heteroaryl,and hydroxy.

The term “halo” represents fluoro, chloro, bromo or iodo. Likewise, theterm “halogen” refers to a fluorine, chlorine, bromine, or iodinesubstituent.

The term “arylalkyl” means an alkyl moiety (as defined above)substituted with one or more aryl moiety (also as defined above). Morepreferred arylalkyl radicals are aryl-C₁₋₃-alkyls. Examples includebenzyl, phenylethyl, and the like.

The term “heteroarylalkyl” means an alkyl moiety (as defined above)substituted with a heteroaryl moiety (also as defined above). Morepreferred heteroarylalkyl radicals are 5- or 6-memberedheteroaryl-C₁₋₃-alkyls. Examples include, but are not limited to,oxazolylmethyl, pyridylethyl and the like.

The term “heterocyclylalkyl” means an alkyl moiety (as defined above)substituted with a heterocyclyl moiety (also defined above). Morepreferred heterocyclylalkyl radicals are 5- or 6-memberedheterocyclyl-Cl_(—)3-alkyls. An example includes, but is not limited to,tetrahydropyranylmethyl.

The term “cycloalkylalkyl” means an alkyl moiety (as defined above)substituted with a cycloalkyl moiety (also defined above). Morepreferred heterocyclyl radicals are 5- or 6-memberedcycloalkyl-C₁₋₃-alkyls. An example includes, but is not limited to,cyclopropylmethyl.

The term “Me” means methyl, “Et” means ethyl, “Bu” means butyl and “Ac”means acetyl.

In general, the various moieties or functional groups of the compoundsof Formula I may be optionally substituted by one or more substituents.Examples of substituents suitable for purposes of this inventioninclude, but are not limited to, halo, alkyl, allyl, alkenyl, alkynyl,heteroalkyl, heteroallyl, heteroalkenyl, heteroalkynyl, alkoxy,heteroalkoxy, G_(n)-cycloalkyl, G_(n)-heterocycloalkyl, G_(n)-OR,G_(n)-NO₂, G_(n)-CN, G_(n)-CO₂R, G_(n)-(C═O)R, G_(n)-O(C═O)R,G_(n)-O-alkyl, G_(n)-OAr, G_(n)-SH, G-SR, G_(n)-SOR, G_(n)-SO²R,G_(n)-S—ArG_(n)-SOAr, G_(n)-O₂Ar, aryl, heteroaryl, G_(n)-Ar,G_(n)-(C═O)NR²R³, G_(n)-NR²R³, G_(n)-NR(C═O)R, G_(n)-SO₂ NR²R³, PO₃H₂,SO₃H₂, where G is alkylene having from 1 to 4 carbons, or alkenylene oralkynylene each having from 2 to 4 carbons; n is zero or 1; R₁, R², andR³ are alkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl,heteroalkenyl, heteroalkynyl, alkoxy, heteroalkoxy, G_(n)-cycloalkyl, orG_(n)-heterocycloalkyl; and Ar is aryl or heteroaryl, wherein saidalkyl, allyl, alkenyl, alkynyl, heteroalkyl, heteroallyl, heteroalkenyl,heteroalkynyl, alkoxy, heteroalkoxy, G_(n)-cycloalkyl,G_(n)-heterocycloalkyl, alkylene, alkenylene, alkynylene, Ar, R¹, R²,and R³ may be further substituted or unsubstituted.

It is to be understood that in instances where two or more radicals areused in succession to define a substituent attached to a structure, thefirst named radical is considered to be terminal and the last namedradical is considered to be attached to the structure in question. Thus,for example, the radical arylalkyl is attached to the structure inquestion by the alkyl group.

The compounds of this invention may possess one or more asymmetriccenters; such compounds can therefore be produced as individual (R)- or(S)-stereoisomers or as mixtures thereof. Unless indicated otherwise,the description or naming of a particular compound in the specificationand claims is intended to include both individual enantiomers andmixtures, racemic or otherwise, thereof. Accordingly, this inventionalso includes racemates and resolved enantiomers, and diastereomerscompounds of the Formula I. Methods for determining the stereochemistryand for the separation of stereoisomers are well known in the art (seediscussion in Chapter 4 of “Advanced Organic Chemistry”, 4th edition J.March, John Wiley and Sons, New York, 1992).

In addition to compounds of the Formula I, the invention also includessolvates, pharmaceutically active metabolites, pharmaceuticallyacceptable prodrugs, and pharmaceutically acceptable salts of suchcompounds.

The term “solvate” refers to an aggregate of a molecule with one or moresolvent molecules.

A “pharmaceutically active metabolite” is a pharmacologically activeproduct produced through metabolism in the body of a specified compoundor salt thereof. Metabolites of a compound may be identified usingroutine techniques known in the art and their activities determinedusing tests such as those described herein.

Compounds of the present invention having functional groups including,but not limited to, free amino, amido, hydroxy or carboxylic groups canbe converted into pharmaceutically acceptable prodrugs. A“pharmaceutically acceptable prodrug” is a compound that may beconverted under physiological conditions or by solvolysis to thespecified compound or to a pharmaceutically acceptable salt of suchcompound. Prodrugs include compounds wherein an amino acid residue, or apolypeptide chain of two or more (e.g., two, three or four) amino acidresidues is covalently joined through an amide or ester bond to a freeamino, hydroxy or carboxylic acid group of compounds of the presentinvention. The amino acid residues include but are not limited to the 20naturally occurring amino acids commonly designated by three lettersymbols and also includes 4-hydroxyproline, hydroxylysine, demosine,isodemosine, 3-methylhistidine, norvaline, beta-alanine,gamma-aminobutyric acid, cirtulline, homocysteine, homoserine, ornithineand methionine sulfone. One preferred prodrug of this invention is acompound of Formula I covalently joined to a phosphate residue. Anotherpreferred prodrug of this invention is a compound of Formula Icovalently joined to a valine residue.

Additional types of prodrugs are also encompassed. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. As anotherexample, compounds of this invention comprising free hydroxy groups maybe derivatized as prodrugs by converting the hydroxy group groupsincluding to a phosphate ester, hemisuccinate, dimethylaminoacetate, orphosphoryloxymethyloxycarbonyl, as outlined in Advanced Drug DeliveryReviews, 1996, 19, 115. Carbamate prodrugs of hydroxy and amino groupsare also included, as are carbonate prodrugs, sulfonate esters andsulfate esters of hydroxy groups. Derivatization of hydroxy groups as(acyloxy)methyl and (acyloxy)ethyl ethers wherein the acyl group may bean alkyl ester, optionally substituted with groups including, but notlimited to, ether, amine and carboxylic acid functionalities, or wherethe acyl group is an amino acid ester as described above, are alsoencompassed. Prodrugs of this type are described in J. Med. Chem., 1996,39, 10. More specific examples include replacement of the hydrogen atomof the alcohol group with a group such as (C₁-C₆)alkanoyloxymethyl,1-((C₁-C₆)alkanoyloxy)ethyl, 1-methyl-1-((C₁-C₆)alkanoyloxy)ethyl,(C₁-C₆)alkoxycarbonyloxymethyl, N—(C₁-C₆)alkoxycarbonylaminomethyl,succinoyl, (C₁-C₆)alkanoyl, α-amino(C₁-C₄)alkanoyl, arylacyl andα-aminoacyl, or α-aminoacyl-α-aminoacyl, where each α-aminoacyl group isindependently selected from the naturally occurring L-amino acids,P(O)(OH)₂, —P(O)(O(C₁-C₆)alkyl)₂ or glycosyl (the radical resulting fromthe removal of a hydroxyl group of the hemiacetal form of acarbohydrate).

Free amines can also be derivatized as amides, sulfonamides orphosphonamides. All of these prodrug moieties may incorporate groupsincluding, but not limited to, ether, amine and carboxylic acidfunctionalities. For example, a prodrug can be formed by the replacementof a hydrogen atom in the amine group with a group such as R-carbonyl,RO-carbonyl, NRR′— carbonyl where R and R′ are each independently(C₁-C₁₀)alkyl, (C₃-C₇)cycloalkyl, benzyl, or R-carbonyl is a naturalα-aminoacyl or natural α-aminoacyl-natural α-aminoacyl, —C(OH)C(O)OYwherein Y is H, (C₁-C₆)alkyl or benzyl, —C(OY₀)Y₁ wherein Y₀ is (C₁-C₄)alkyl and Y₁ is (C₁-C₆)alkyl, carboxy(C₁-C₆)alkyl, amino(C₁-C₄)alkyl ormono-N— or di-N,N—(C₁-C₆)alkylaminoalkyl, —C(Y₂)Y₃ wherein Y₂ is H ormethyl and Y₃ is mono-N— or di-N,N—(C₁-C₆)alkylamino, morpholino,piperidin-1-yl or pyrrolidin-1-yl.

A “pharmaceutically acceptable salt” is a salt that retains thebiological effectiveness of the free acids and bases of the specifiedcompound and that is not biologically or otherwise undesirable. Acompound of the invention may possess a sufficiently acidic, asufficiently basic, or both functional groups, and accordingly reactwith any of a number of inorganic or organic bases, and inorganic andorganic acids, to form a pharmaceutically acceptable sale. Examples ofpharmaceutically acceptable salts include those salts prepared byreaction of the compounds of the present invention with a mineral ororganic acid or an inorganic base, such salts including sulfates,pyrosulfates, bisulfates, sulfites, bisulfites, phosphates,monohydrogenphosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates, propionates,decanoates, caprylates, acrylates, formates, isobutyrates, caproates,heptanoates, propiolates, oxalates, malonates, succinates, suberates,sebacates, fumarates, maleates, butyn-1,4-dioates, hexyne-1,6-dioates,benzoates, chlorobenzoates, methylbenzoates, dinitromenzoates,hydroxybenzoates, methoxybenzoates, phthalates, sulfonates,xylenesulfonates, pheylacetates, phenylpropionates, phenylbutyrates,citrates, lactates, γ-hydroxybutyrates, glycollates, tartrates,methanesulfonates, propanesulfonates, naphthalene-1-sulfonates,naphthalene-2-sulfonates, and mandelates.

If the inventive compound is a base, the desired pharmaceuticallyacceptable salt may be prepared by any suitable method available in theart, for example, treatment of the free base with an inorganic acid,such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid and the like, or with an organic acid, such as aceticacid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonicacid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, apyranosidyl acid such as glucuronic acid or galacturonic acid, analphahydroxy acid such as citric acid or tartaric acid, an amino acidsuch as aspartic acid or glutamic acid, an aromatic acid such as benzoicacid or cinnamic acid, a sulfonic acid such as p-toluenesulfonic acid orethanesulfonic acid, or the like.

If the inventive compound is an acid, the desired pharmaceuticallyacceptable salt may be prepared by any suitable method, for example,treatment of the free acid with an inorganic or organic base, such as anamine (primary, secondary or tertiary), an alkali metal hydroxide oralkaline earth metal hydroxide, or the like. Illustrative examples ofsuitable salts include, but are not limited to, organic salts derivedfrom amino acids, such as glycine and arginine, ammonia, primary,secondary, and tertiary amines, and cyclic amines, such as piperidine,morpholine and piperazine, and inorganic salts derived from sodium,calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminumand lithium.

The inventive compounds may be prepared using the reaction routes andsynthesis schemes as described herein, employing the techniquesavailable in the art using starting materials that are readilyavailable.

Therapeutic Aspects of the Invention

The invention also provides a pharmaceutical composition for thetreatment of a hyperproliferative disorder in a mammal which comprises atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, prodrug, metabolite or hydratethereof, and a pharmaceutically acceptable carrier. In one embodiment,said pharmaceutical composition is for the treatment of cancer such asskin, brain, lung, squamous cell, bladder, gastric, pancreatic, breast,head, neck, renal, kidney, ovarian, prostate, colorectal, esophageal,testicular, gynecological, cardiac, liver, bone, meninges, spinal cord,blood, skin, adrenal or thyroid cancer. In another embodiment, saidpharmaceutical composition is for the treatment of a non-canceroushyperproliferative disorder such as benign hyperplasia of the skin(e.g., psoriasis), restenosis, or prostate (e.g., benign prostatichypertrophy (BPH)).

The invention also relates to a method for the treatment of ahyperproliferative disorder in a mammal that comprises administering tosaid mammal a therapeutically effective amount of a compound of thepresent invention, or a pharmaceutically acceptable salt, prodrug orhydrate thereof, in combination with an anti-tumor agent selected fromthe group consisting of mitotic inhibitors, alkylating agents,anti-metabolites, intercalating antibiotics, growth factor inhibitors,cell cycle inhibitors, enzyme inhibitors, topoisomerase inhibitors,biological response modifiers, anti-hormones, angiogenesis inhibitors,and anti-androgens.

The invention also relates to a method of treating pancreatitis orkidney disease in a mammal that comprises administering to said mammal atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, prodrug or hydrate thereof.

The invention also relates to a method of preventing blastocyteimplantation in a mammal that comprises administering to said mammal atherapeutically effective amount of a compound of the present invention,or a pharmaceutically acceptable salt, prodrug or hydrate thereof.

The invention also relates to a method of treating diseases related tovasculogenesis or angiogenesis in a mammal that comprises administeringto said mammal a therapeutically effective amount of a compound of thepresent invention, or a pharmaceutically acceptable salt, prodrug orhydrate thereof. In one embodiment, said method is for treating adisease selected from the group consisting of tumor angiogenesis,chronic inflammatory disease such as rheumatoid arthritis,atherosclerosis, inflammatory bowel disease, skin diseases such aspsoriasis, excema, and scleroderma, diabetes, diabetic retinopathy,retinopathy of prematurity, age-related macular degeneration,hemangioma, glioma, melanoma, Kaposi's sarcoma and ovarian, breast,lung, pancreatic, prostate, colon and epidermoid cancer.

The invention also relates to a pharmaceutical composition for treatinga disease or condition related to inflammatory disease, autoimmunedisease, destructive bone disorders, proliferative disorders, infectiousdisease, viral disease, fibrotic disease or neurodegenerative disease ina mammal which comprises a therapeutically effective amount of acompound of the present invention, or a pharmaceutically acceptablesalt, prodrug or hydrate thereof, and a pharmaceutically acceptablecarrier. Examples of the above diseases and/or conditions include but isnot limited to rheumatoid arthritis, atherosclerosis, inflammatory boweldisease, skin diseases such as psoriasis, eczema, and scleroderma,diabetes and diabetic complications, diabetic retinopathy, retinopathyof prematurity, age-related macular degeneration, hemangioma, chronicobstructive pulmonary disease, idiopathic pulmonary fibrosis, allergicresponses including asthma allergic rhinitis and atopic dermatitis,renal disease and renal failure, polycystic kidney disease, acutecoronary syndrome, congestive heart failure, osteoarthritis,neurofibromatosis, organ transplant rejection, cachexia and pain.

Further provided is a compound of Formula I for use as a medicament inthe treatment of the diseases and conditions described above in awarm-blooded animal, preferably a mammal, more preferably a human,suffering from such disorder. Also provided is the use of a compound ofFormula I in the preparation of a medicament for the treatment of thediseases and conditions described above in a warm-blooded animal,preferably a mammal, more preferably a human, suffering from suchdisorder.

Patients that can be treated with compounds of the present invention, orpharmaceutically acceptable salts, prodrugs and hydrates of saidcompounds, according to the methods of this invention include, forexample, patients that have been diagnosed as having psoriasis,restenosis, atherosclerosis, BPH, lung cancer, bone cancer, CMML,pancreatic cancer, skin cancer, cancer of the head and neck, cutaneousor intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer,cancer of the anal region, stomach cancer, colon cancer, breast cancer,testicular, gynecologic tumors (e.g., uterine sarcomas, carcinoma of thefallopian tubes, carcinoma of the endometrium, carcinoma of the cervix,carcinoma of the vagina or carcinoma of the vulva), Hodgkin's disease,cancer of the esophagus, cancer of the small intestine, cancer of theendocrine system (e.g., cancer of the thyroid, parathyroid or adrenalglands), sarcomas of soft tissues, cancer of the urethra, cancer of thepenis, prostate cancer, chronic or acute leukemia, solid tumors ofchildhood, lymphocytic lymphomas, cancer of the bladder, cancer of thekidney or ureter (e.g., renal cell carcinoma, carcinoma of the renalpelvis), or neoplasms of the central nervous system (e.g., primary CNSlymphona, spinal axis tumors, brain stem gliomas or pituitary adenomas).

This invention also relates to a pharmaceutical composition forinhibiting abnormal cell growth in a mammal which comprises an amount ofa compound of the present invention, or a pharmaceutically acceptablesalt or solvate or prodrug thereof, in combination with an amount of achemotherapeutic, wherein the amounts of the compound, salt, solvate, orprodrug, and of the chemotherapeutic are together effective ininhibiting abnormal cell growth. Many chemotherapeutics are presentlyknown in the art. In one embodiment, the chemotherapeutic is selectedfrom the group consisting of mitotic inhibitors, alkylating agents,anti-metabolites, antitumor antibiotics, growth factor inhibitors, cellcycle inhibitors, enzymes, topoisomerase inhibitors, biological responsemodifiers, anti-hormones, angiogenesis inhibitors, and anti-androgens.

This invention further relates to a method for inhibiting abnormal cellgrowth in a mammal or treating a hyperproliferative disorder whichmethod comprises administering to the mammal an amount of a compound ofthe present invention, or a pharmaceutically acceptable salt or solvateor prodrug thereof, in combination with radiation therapy, wherein theamounts of the compound, salt, solvate, or prodrug, is in combinationwith the radiation therapy effective in inhibiting abnormal cell growthor treating the hyperproliferative disorder in the mammal. Techniquesfor administering radiation therapy are known in the art, and thesetechniques can be used in the combination therapy described herein. Theadministration of the compound of the invention in this combinationtherapy can be determined as described herein.

It is believed that the compounds of the present invention can renderabnormal cells more sensitive to treatment with radiation for purposesof killing and/or inhibiting the growth of such cells. Accordingly, thisinvention further relates to a method for sensitizing abnormal cells ina mammal to treatment with radiation which comprises administering tothe mammal an amount of a compound of the present invention orpharmaceutically acceptable salt or solvate or prodrug thereof, whichamount is effective is sensitizing abnormal cells to treatment withradiation. The amount of the compound, salt, or solvate in this methodcan be determined according to the means for ascertaining effectiveamounts of such compounds described herein.

Compounds and methods of this invention may also be used to treat otherdiseases and conditions (e.g., inflammatory disease), includingrheumatoid arthritis, osteoarthritis, endometriosis, atherosclerosis,vein graft stenosis, peri-anastomatic prosthetic graft stenosis,prostate hyperplasia, chronic obstructive pulmonary disease, psoriasis,inhibition of neurological damage due to tissue repair, scar tissueformation (and can aid in wound healing), multiple sclerosis,inflammatory bowel disease, infections, particularly bacterial, viral,retroviral or parasitic infections (by increasing apoptosis), pulmonarydisease, neoplasm, Parkinson's disease, transplant rejection (as animmunosuppressant), macular degeneration and septic shock.

Therapeutically effective amounts of the compounds of the invention maybe used to treat diseases mediated by modulation or regulation of AKTprotein kinases, tyrosine kinases, additional serine/threonine kinases,and/or dual specificity kinases. An “effective amount” is intended tomean that amount of compound that, when administered to a mammal in needof such treatment, is sufficient to effect treatment for a diseasemediated by the activity of one or more AKT protein kinases, tyrosinekinases, additional serine/threonine kinases, and/or dual specificitykinases. Thus, for example, a therapeutically effective amount of acompound selected from Formula I or a salt, active metabolite or prodrugthereof, is a quantity sufficient to modulate, regulate, or inhibit theactivity of one or more AKT protein kinases, tyrosine kinases,additional serine/threonine kinases, and/or dual specificity kinasessuch that a disease condition which is mediated by that activity isreduced or alleviated.

The terms “abnormal cell growth” and “hyperproliferative disorder” areused interchangeably in this application.

“Abnormal cell growth,” as used herein, unless otherwise indicated,refers to cell growth that is independent of normal regulatorymechanisms (e.g., loss of contact inhibition). This includes, forexample, the abnormal growth of: (1) tumor cells (tumors) thatproliferate by expressing a mutated tyrosine kinase or overexpression ofa receptor tyrosine kinase; (2) benign and malignant cells of otherproliferative diseases in which aberrant tyrosine kinase activationoccurs; (3) any tumors that proliferate by receptor tyrosine kinases;(4) any tumors that proliferate by aberrant serine/threonine kinaseactivation; and (5) benign and malignant cells of other proliferativediseases in which aberrant serine/theroine kinase activation occurs.

The amount of a given agent that will correspond to such an amount willvary depending upon factors such as the particular compound, diseasecondition and its severity, the identity (e.g., weight) of the mammal inneed of treatment, but can nevertheless be routinely determined by oneskilled in the art. “Treating” is intended to mean at least themitigation of a disease condition in a mammal, such as a human, that isaffected, at least in part, by the activity of one or more AKT proteinkinases, tyrosine kinases, additional serine/threonine kinases, and/ordual specificity kinases, and includes, but is not limited to,preventing the disease condition from occurring in a mammal,particularly when the mammal is found to be predisposed to having thedisease condition but has not yet been diagnosed as having it;modulating and/or inhibiting the disease condition; and/or alleviatingthe disease condition.

In order to use a compound of the Formula I or a pharmaceuticallyacceptable salt, solvate, metabolite or prodrug thereof, for thetherapeutic treatment (including prophylactic treatment) of mammalsincluding humans, it is normally formulated in accordance with standardpharmaceutical practice as a pharmaceutical composition. According tothis aspect of the invention there is provided a pharmaceuticalcomposition that comprises a compound of the Formula I, or apharmaceutically acceptable salt, solvate, metabolite or prodrugthereof, as defined hereinbefore in association with a pharmaceuticallyacceptable diluent or carrier.

To prepare the pharmaceutical compositions according to this invention,a therapeutically or prophylactically effective amount of a compound ofFormula I or pharmaceutically acceptable salt, solvate, metabolite orprodrug thereof (alone or together with an additional therapeutic agentas disclosed herein) is preferably intimately admixed with apharmaceutically acceptable carrier according to conventionalpharmaceutical compounding techniques to produce a dose. A carrier maytake a wide variety of forms depending on the form of preparationdesired for administration, e.g., oral or parenteral. Examples ofsuitable carriers include any and all solvents, dispersion media,adjuvants, coatings, antibacterial and antifungal agents, isotonic andabsorption delaying agents, sweeteners, stabilizers (to promote longterm storage), emulsifiers, binding agents, thickening agents, salts,preservatives, solvents, dispersion media, coatings, antibacterial andantifungal agents, isotonic and absorption delaying agents, flavoringagents, and miscellaneous materials such as buffers and absorbents thatmay be needed in order to prepare a particular therapeutic composition.The use of such media and agents with pharmaceutically active substancesis well known in the art. Except insofar as any conventional media oragent is incompatible with a compound of Formula I, its use in thetherapeutic compositions and preparations is contemplated. Supplementaryactive ingredients can also be incorporated into the compositions andpreparations as described herein.

The compositions of the invention may be in a form suitable for oral use(for example as tablets, lozenges, hard or soft capsules, aqueous oroily suspensions, emulsions, dispersible powders or granules, syrups orelixirs), for topical use (for example as creams, ointments, gels, oraqueous or oily solutions or suspensions), for administration byinhalation (for example as a finely divided powder or a liquid aerosol),for administration by insuffiation (for example as a finely dividedpowder) or for parenteral administration (for example as a sterileaqueous or oily solution for intravenous, subcutaneous, or intramusculardosing or as a suppository for rectal dosing). For example, compositionsintended for oral use may contain one or more coloring, sweetening,flavoring and/or preservative agents.

Suitable pharmaceutically-acceptable excipients for a tablet formulationinclude, for example, inert diluents such as lactose, sodium carbonate,calcium phosphate or calcium carbonate, granulating and disintegratingagents such as corn starch or algenic acid; binding agents such asstarch; lubricating agents such as magnesium stearate, stearic acid ortalc; preservative agents such as ethyl or propyl p-hydroxybenzoate, andanti-oxidants, such as ascorbic acid. Tablet formulations may beuncoated or coated either to modify their disintegration and thesubsequent absorption of the active ingredient within thegastrointestinal tract, or to improve their stability and/or appearance,in either case, using conventional coating agents and procedures wellknown in the art.

Compositions for oral use may be in the form of hard gelatin capsules inwhich the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules in which the active ingredient is mixed with water oran oil such as peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions generally contain the active ingredient in finelypowdered form together with one or more suspending agents, such assodium carboxymethylcellulose, methylcellulose,hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone,gum tragacanth and gum acacia; dispersing or wetting agents such aslecithin or condensation products of an alkylene oxide with fatty acids(for example polyoxethylene stearate), or condensation products ofethylene oxide with long chain aliphatic alcohols, for exampleheptadecaethyleneoxycetanol, or condensation products of ethylene oxidewith partial esters derived from fatty acids and a hexitol such aspolyoxyethylene sorbitol monooleate, or condensation products ofethylene oxide with partial esters derived from fatty acids and hexitolanhydrides, for example polyethylene sorbitan monooleate. The aqueoussuspensions may also contain one or more preservatives (such as ethyl orpropyl p-hydroxybenzoate, anti-oxidants (such as ascorbic acid),coloring agents, flavoring agents, and/or sweetening agents (such assucrose, saccharine or aspartame).

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil (such as arachis oil, olive oil, sesame oil orcoconut oil) or in a mineral oil (such as liquid paraffin). The oilysuspensions may also contain a thickening agent such as beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set outabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water generally contain the activeingredient together with a dispersing or wetting agent, suspendingagent, and one or more preservatives. Suitable dispersing or wettingagents and suspending agents are exemplified by those already mentionedabove. Additional excipients such as sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, suchas olive oil or arachis oil, or a mineral oil such as liquid paraffin,or a mixture of any of these. Suitable emulsifying agents may be, forexample, naturally-occurring gums such as gum acacia or gum tragacanth,naturally-occurring phosphatides such as soya bean, lecithin, an estersor partial esters derived from fatty acids and hexitol anhydrides (forexample sorbitan monooleate) and condensation products of the saidpartial esters with ethylene oxide such as polyoxyethylene sorbitanmonooleate. The emulsions may also contain sweetening, flavoring andpreservative agents.

Syrups and elixirs may be formulated with sweetening agents such asglycerol, propylene glycol, sorbitol, aspartame or sucrose, and may alsocontain a demulcent, preservative, flavoring and/or coloring agent.

The pharmaceutical compositions may also be in the form of a sterileinjectable aqueous or oily suspension, which may be formulated accordingto known procedures using one or more of the appropriate dispersing orwetting agents and suspending agents, which have been mentioned above. Asterile injectable preparation may also be a sterile injectable solutionor suspension in a non-toxic parenterally-acceptable diluent or solvent,for example a solution in 1,3-butanediol.

Suppository formulations may be prepared by mixing the active ingredientwith a suitable non-irritating excipient, which is solid at ordinarytemperature s but liquid at the rectal temperature and will thereforemelt in the rectum to release the drug. Suitable excipients include, forexample, cocoa butter and polyethylene glycols.

Topical formulations, such as creams, ointments, gels and aqueous oroily solutions or suspensions, may generally be obtained by formulatingan active ingredient with a conventional, topically acceptable, vehicleor diluent using conventional procedures well known in the art.

Compositions for administration by insufflation may be in the form of afinely divided powder containing particles of average diameter of, forexample, 30 μm or much less, the powder itself comprising either activeingredient alone or diluted with one or more physiologically acceptablecarriers such as lactose. The powder for insufflation is thenconveniently retained in a capsule containing, for example, 1 to 50 mgof active ingredient for use with a turbo-inhaler device, such as isused for insufflation of the known agent sodium cromoglycate.

Compositions for administration by inhalation may be in the form of aconventional pressurized aerosol arranged to dispense the activeingredient either as an aerosol containing finely divided solid orliquid droplets. Conventional aerosol propellants such as volatilefluorinated hydrocarbons or hydrocarbons may be used and the aerosoldevice is conveniently arranged to dispense a metered quantity of activeingredient.

For further information on formulations, see Chapter 25.2 in Volume 5 ofComprehensive Medicinal Chemistry (Corwin Hansch; Chairman of EditorialBoard), Pergamon Press 1990, which is specifically incorporated hereinby reference.

The amount of a compound of this invention that is combined with one ormore excipients to produce a single dosage form will necessarily varydepending upon the host treated and the particular route ofadministration. For example, a formulation intended for oraladministration to humans may contain, for example, from 0.5 mg to 2 g ofactive agent compounded with an appropriate and convenient amount ofexcipients, which may vary from about 5 to about 98 percent by weight ofthe total composition. Dosage unit forms will generally contain about 1mg to about 500 mg of an active ingredient. For further information onroutes of administration and dosage regimes, see Chapter 25.3 in Volume5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman ofEditorial Board), Pergamon Press 1990, which is specificallyincorporated herein by reference.

The size of the dose for therapeutic or prophylactic purposes of acompound of Formula I will naturally vary according to the nature andseverity of the conditions, the age and sex of the animal or patient andthe route of administration, according to well known principles ofmedicine.

The compounds of this invention may be used alone in combination withother drugs and therapies used in the treatment of disease states whichwould benefit from the inhibition of MEK. Such treatment may involve, inaddition to the compounds of the invention, conventional surgery orradiotherapy or chemotherapy. Such chemotherapy may include one or moreof the following categories of anti-tumor agents:

(i) antiproliferative/antineoplastic drugs and combinations thereof, asused in medical oncology, such as alkylating agents (for example,cis-platin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan,chlorambucil, busulphan and nitorsoureas); anti-metabolites (forexample, antifolates such as such as fluoropyrimidines like5-fluorouracil and tegafur, raltitrexed, methotrexate, cytosinearabinside, hydroxyurea, or, one of the preferred anti-metabolitesdisclosed in European Patent Application No. 239362 such asN-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamicacid); antitumor antibiotics (for example, anthracyclines likeadriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin,mitomycin-C, dactinomycin and mithramycin); antimitotic agents (forexample, vinca alkaloids like vincristine, vinblastine, vindesine andvinorelbine and taxoids like taxol and taxotere); and topoisomeraseinhibitors (for example epipodophyllotoxins like eptoposide andteniposide, amsacrine, topotecan and campothecin):

(ii) cytostatic agents such as antiestrogens (for example, tamoxifen,toremifene, raloxifene, droloxifene and iodoxyfene), estrogen receptordown regulators (for example, fulvestrant), antiandrogens (for example,bicalutamide, flutamide, nilutamide, cyproterone acetate and Casodex™(4′-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3′-(trifluoromethyl)propionanilide)),LHRH antagonists or LHRH agonists (for example, goserelin, leuporelinand buserelin), progestogens (for example, megestrol acetate), aromataseinhibitors (for example, anastrozole, letrozole, vorazole andexemestane) and inhibitors of 5α-reductase such as finasteride;

(iii) agents which inhibit cancer cell invasion (for example,metalloproteinase inhibitors like marimastat and inhibitors of urokinaseplasminogne activator receptor function);

(iv) inhibitors of growth factor function like growth factor antibodies,growth factor receptor antibodies (for example, the anti-erbB2 antibodytrastumuzab [Herceptin™] and the anti-erbB1 antibody cetuximab [C225]),farnesyl transferase inhibitors, tyrosine kinase inhibitors andserine-threonine kinase inhibitors (for example, inhibitors of theepidermal growth factor family tyrosine kinases such asN-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine(gefitinib, AZD1839),N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine(erlotinib, OSI-774) and6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)quinazolin-4-amine(CI 1033)), inhibitors of the platelet-derived growth factor family andinhibitors of the hepatocyte growth factor family;

(v) antiangiogenic agents such as those which inhibit the effects ofvascular endothelial growth factor (for example, the anti-vascularendothelial cell growth factor antibody bevacizumab [Avastin™],compounds such as those disclosed in International Patent ApplicationsWO 97/22596, WO 97/30035, WO 97/32856, and WO 98/13354) and compoundsthat work by other mechanisms (for example, linomide, inhibitors ofintegrin αv(33 function, MMP inhibitors, COX-2 inhibitors andangiostatin);

(vi) vascular damaging agents such as Combretastatin A4 and compoundsdisclosed in International Patent Applications WO 99/02166, WO 00/40529,WO 00/41669, WO 01/92224, WO 02/04434, and WO 02/08213;

(vii) antisense therapies (for example, those which are directed to thetargets listed above such as ISIS 2503, and anti-ras antisense);

(viii) gene therapy approaches, including for example GVAX™, approachesto replace aberrant genes such as aberrant p53 or aberrant BRCA1 orBRCA2, GDEPT (gene-directed enzyme prodrug therapy) approaches such asthose using cytosine deaminase, thymidine kinase or a bacterialnitroreductase enzyme and approaches to increase patient tolerance tochemotherapy or radiotherapy such as multi-drug resistance gene therapy;

(ix) interferon; and

(x) immunotherapy approaches, including for example ex-vivo and in-vivoapproaches to increase the immunogenicity of patient tumor cells, suchas transfection with cytokines such as interleukin 2, interleukin 4 orgranulocyte-macrophage colony stimulating factor, approaches to decreaseT-cell anergy, approaches to using transfected immune cells such ascytokine-transfected dendritic cells, approaches usingcytokine-transfected tumor cell lines and approaches usinganti-idiotypic antibodies.

Such conjoint treatment may be achieved by way of the simultaneous,sequential or separate dosing of the individual components of treatment.Such combination products employ the compounds of this invention withinthe dose range described hereinbefore and the other pharmaceuticallyactive agent within its approved dose range.

According to this aspect of the invention there is provided apharmaceutical product comprising a compound of Formula I as definedhereinbefore and an additional anit-tumor agent as definged hereinbeforefor the conjoint treatment of cancer.

Although the compounds of Formula I are primarily of value astherapeutic agents for use in warm-blooded animals (including man), theyare also useful whenever it is required to control AKT protein kinases,tyrosine kinases, additional serine/threonine kinases, and/or dualspecificity kinases. Thus, they are useful as pharmacological standardsfor use in the development of new biological tests and in the search fornew pharmacological agents.

The activity of the compounds of this invention may be assayed for AKTprotein kinases, tyrosine kinases, additional serine/threonine kinases,and/or dual specificity kinases in vitro, in vivo, or in a cell line. Invitro assays include assays that determine inhibition of the kinaseactivity. Alternate in vitro assays quantitate the ability of theinhibitor to bind to kinases and may be measured either byradiolabelling the inhibitor prior to binding, isolating theinhibitor/kinase complex and determining the amount of radiolabel bound,or by running a competition experiment where new inhibitors areincubated with known radioligands. These and other useful in vitro andcell culture assays are well known to those of skill in the art.

Although the invention has been described and illustrated with a certaindegree of particularity, it is understood that the present disclosurehas been made only by way of example, and that numerous changes in thecombination and arrangement of parts can be resorted to by those skilledin the art without departing from the spirit and scope of the invention,as hereinafter claimed.

The compounds of the present invention may be prepared in a number ofways well known to one skilled in the art of organic synthesis. Thecompounds of the present invention can be synthesized using the methodsdescribed below, together with synthetic methods known in the art ofsynthetic organic chemistry, or variations thereon as appreciated bythose skilled in the art. Preferred methods include, but are not limitedto, those described below.

The novel compounds of the present invention may be prepared using thereactions and techniques described in this section. The reactions areperformed in solvents appropriate to the reagents and materials employedand are suitable for the transformations being effected. Also, in thedescription of the synthetic methods described below, it is to beunderstood that all proposed reaction conditions, including choice ofsolvent, reaction atmosphere, reaction temperature, duration of theexperiment and workup procedures, are chosen to be the conditionsstandard for that reaction, which should be readily recognized by oneskilled in the art. It is understood by one skilled in the art oforganic synthesis that the functionality present on various portions ofthe molecule must be compatible with the reagents and reactionsproposed. Such restrictions to the substituents that are compatible withthe reaction conditions will be readily apparent to one skilled in theart and alternate methods must then be used.

The preparation of compounds of the present invention may be carried outin a convergent or sequential synthetic manner. The skills required inpreparation and purification of such compounds and the intermediatesleading to these compounds are known to those in the art. Purificationprocedures include, but are not limited to, normal or reverse phasechromatography, crystallization, and distillation.

An illustration of the preparation of compounds (8), (9), (10) and (11)of the present invention is shown in FIG. 1. The synthesis starts withthe preparation of a substituted quinazolinone (3) made by, for example,by the condensation of a corresponding substituted aryl amino acid (1)and a corresponding substituted amide (2) (see, for example, LeMahieu,et al, J. Med. Chem., 1983, 26, 420-5 and references cited therein).Introduction of a leaving group into quinazolinone (3) may beaccomplished by treatment with a halogenating agent (for example POCl₃)to give the chlorinated quinazoline (4.) The halogen leaving group isthen displaced with substituted and protected piperazine (5) (e.g., Boc,but any suitable protecting group may be used; see, T. W. Greene et al.,‘Protective groups in organic synthesis’, John Wiley and Sons, 1999,3^(rd) Ed., pp. 494-653). The piperazine (5) may be introduced tochlorinated quinazoline (4) either neat or in the presence of base. Thepiperazine protecting group may then be removed by known methods (see,Greene et al, supra) (6).

Substitution of the piperazine secondary amine in quinazolineintermediate (6) may be accomplished using a variety of electrophilesand reaction conditions. For example, the piperazine may be acylated bya suitably N-substituted or protected amino acid (e.g., Boc, etc.) whichmay be introduced using a variety of standard peptide couplingprocedures under both solution phase and solid phase conditions, toproduce a product such as compound (8). For representative examples, seeMiklos Bodanszky, ‘Principles of Peptide Synthesis,’ Springer-Verlag,1993, 2nd Ed., and C. Najera, Synlett, 2002, 9, 1388-1403. As above (andif protected) the N-protected amino acid unit may then be deprotectedusing representative procedures (e.g., using acid on a Boc-group; Greeneet al., supra), and then manipulated as desired according to proceduresappreciated by those skilled in the art.

Compounds of the present invention similar to compound (8) may be aprepared from quinazoline intermediate (6) by acylation with a naturalor an ‘unnatural’ amino acid (7). The preparation of ‘unnatural’ aminoacids is also well known to those skilled in the art, and their use isincluded in the present invention (for representative reviews, see, C.Najera, Synlett, 2002, 9, 1388-1403, and J.-A. Ma, Angew. Chemie, Int.Ed., 2003, 42, 4290-4299, and references therein).

Alternatively, the piperazine (6) may be acylated with an acid or acidhalide in the presence of base to generate a substituted amine (10).Additionally, a substituted tertiary amine (11) can be prepared bytreating piperazine (6) with an appropriate aldehyde (or surrogate) inthe presence of a reducing agent (e.g., sodium cyanoborohydride). Thepiperazine (6) can also be treated with an epoxide to give the aminoalcohol (9.) All functional groups may be further manipulated understandard conditions (e.g., reductions, alkylations, oxidations,palladium or nickel mediated couplings, etc.) to further functionalizeeach compound.

The compounds described in FIG. 1 may be prepared either as either theracemate, or as a single enantiomer (for example, using anenantiomerically pure amino acid (7.)) If prepared as the racemate, thecorresponding enantiomers may be isolated by separation of the racemicmixture of compound on a chiral stationary phase column utilizing normalor reverse phase HPLC techniques. Alternatively, a diastereomericmixture of compound (8) can be prepared by treatment of racemic compound(8) with an appropriate chiral acid (or suitably activated derivative),for example dibenzoyl tartrate or the like (see, for example, Kinbara,K., et. al., J. Chem. Soc., Perkin Trans. 2, 1996, 2615; and Tomori, H.,et. al., Bull. Chem. Soc. Jpn., 1996, 3581). The diastereomers wouldthen be separated by traditional techniques (i.e. silica chromatography,crystallization, HPLC, etc) followed by removal of the chiral auxiliaryto afford enantiomerically pure compound (8.)

Other compounds of the present invention can be prepared using thealternatively substituted and functionalised acids, amino acids, hydroxyacids and variants thereof described in FIGS. 2-15. For example,compound (14) of this invention may be prepared as shown in FIG. 2. Thehydroxyl group of substituted α-hydroxy benzyl ester (12) is protectedwith an appropriate protecting group (such as acetate) to give compound13. The benzyl ester is then converted to the corresponding carboxylicacid (for example by hydrogenolysis) to give compound (14).

FIG. 3 shows the preparation of compound (18). Substituted phenylboronic acid (15), glyoxylic acid (16), and a chiral or achiralmono-protected (using the Boc protecting group, for example) diamine(17) (such as 3-Boc-aminopyrrolidine) are combined in an appropriatesolvent such as 1,2-dichloroethane and stirred at elevated temperatureto provide carboxylic acids (18).

FIG. 4 shows the preparation of compound (22). 2-(2-Aminoethoxy)ethanol(19) is protected with an appropriate amine protecting group (such asBoc), and the hydroxyl group is oxidized to the carboxylic acid toprovide intermediate (20). The acid in compound (20) is then convertedto an ester using an appropriate base (such as K₂CO₃) and alkyl halideto furnish intermediate compound (21). Enolization of intermediatecompound (21) is accomplished with strong base (such as LDA or LHMDS),followed by addition of a substituted benzyl halide yields an alkylatedester, which is then converted by basic hydrolysis to the correspondingacid (22).

Compound (28) may be prepared as shown in FIG. 5. The substituted phenylcarboxylic acid (23) is transformed to the appropriate ester (24) underacidic (mineral acid, R²OH) or basic (K₂CO₃, R²X) conditions.Enolization of ester (24) is accomplished with strong base (such asLDA), and addition of a haloacetate ester (for example tert-butylbromoacetate) provides intermediate compound (25). Selective esterdeprotection is performed by treating compound (25) with acid (such asTFA) to provide carboxylic acid (26). The carboxylic acid (26) isconverted to an acyl azide (using diphenylphosphoryl azide, forexample), which is then transformed to the correspondingcarbamate-protected amine by heating in an appropriate alcohol solvent(tert-butyl alcohol, for example) in the presence or absence of a Lewisacid (such as SnCL4) to provide compound (27). The carboxylic acid esteris then converted to the corresponding acid by hydrolysis under basicconditions (such as aq. LiOH in THF) to afford compound (28).

FIG. 6 shows the preparation of carboxylic acid (31). Lactam (29) isenolized with strong base (such as LDA/LiBr or LHMDS), and addition of asubstituted benzyl halide furnishes alkylated intermediate compound(30). The lactam is then opened under basic conditions (such as aq.LiOH, THF) to furnish carboxylic acid (31).

Compound (36) may be prepared as shown in FIG. 7. Condensation of anappropriately substituted benzaldehyde with ethyl cyanoacetate providescompounds of structure (32). Treatment with a reducing agent such asNaBH₄ gives the saturated compound (33), which is followed bycobalt-mediated hydride reduction to give compound (34). The amine canthen be protected and the ester saponified to give compound (35).Coupling with a piperazine can be accomplished using (for example) EDCIor PyBrop, followed by deprotection to give final compound (36).

FIG. 8 shows the preparation of amino acid (40). Compound (39) can beprepared by condensation of benzaldehydes with ethyl cyanoacetatefollowed by catalytic hydrogenation according to the proceduresdescribed by Lee, J. et al. (1999), 3060-3065. Compound (39) can beconverted to compound amino acid (40) by protection of the primary aminefollowed by saponification under basic condition (for example, aqueousLiOH solution).

An alternate approach to amino acid (40) is shown in FIG. 9. Compound(41), where Pg is an appropriate protecting group (for example, Boc),can be treated with a variety of organometallic agents such as LDA in asuitable solvent such as THF or ether at low temperature s to generate adianion intermediate, which can be quenched by suitable amount of benzylhalides to afford the intermediate compound (42). Saponification underbasic conditions such as aqueous LiOH solution furnishes the desiredproduct (40).

FIG. 10 summarizes a synthesis of amino alcohols (46) from compound(45). Compound (45) may be prepared from compound (44) by a sequence ofdeprotonation, alkylation and saponification as described in FIG. 9.

Preparation of compound (50) is shown in FIG. 11. Phenyl acetic acidderivative (47) can be deprotonated by treatment with a suitableorganometallic agent such as LDA in a suitable solvent such as THF orether at low temperature s, and then reacted with compound (48), where Xis a suitable leaving group (for example Br, Cl) and Pg is anappropriate protecting group (for example, Boc or Ts), to yieldintermediate compound (49) (Ho-sam A. et al. (1997) J. Med. Chem., 40,2196; Ohkanda et al. (2004), J. Med. Chem., 47, 432). Saponification ofcompound (49) under basic conditions (for example, aqueous LiOHsolution) gives acid (50).

Compound (55) may be prepared as shown in FIG. 12. Esterification of theappropriately substituted and commercially available acid (51) with analcohol affords the desired ester (52). Treatment of ester (52) withappropriate base and electrophile (e.g., acrylate, etc. (53)) followedby ester cleavage with acid (see, T. W. Greene et al., supra) affordsintermediate compound (54). Introduction of azide (affords acyl-azide)with activating reagent followed by heating affects rearrangement ofacid (54) to the requisite N-protected amino-ester intermediate (forexample Boc, but any suitable protecting group may be used with theappropriate alcohol solvent; see, Greene et al., supra. Treatment ofester intermediate with hydroxide base affords the N-protected aminoacid (55).

Compound (58) may be prepared as shown in FIG. 13. Michael addition ofphenylacetic acid ethyl esters with tert-butyl acrylate using catalyticbase such as potassium tert-butoxide followed by acid hydrolysis of thetert-butyl ester provides compound (56). Curtius rearrangement usingdiphenylphosphorylazide followed by saponification of the ester givescompound (57). Coupling with a piperazine can be accomplished using EDCIor PyBrop, followed by deprotection of the Boc group to give finalcompound (58).

FIG. 14 shows the preparation of compound (61). Alkylation ofphenylacetic acid ethyl esters with α-bromoacetate tert-butyl esterusing a base such as lithium bis(trimethylsilyl)amide provides compound(59). The remainder of the sequence is as that described in FIG. 13 toprovide compound (61).

Compound (71) may be prepared as shown in FIG. 15. The displacement of4-chloroquinazoline with ethyl isonipecotate followed by saponificationof the ester gives intermediate (68). Treatment with a halogenatingreagent such as thionyl chloride or oxalyl chloride provides acidchloride (69). Reductive amination of an appropriately substitutedbenzaldehyde with N-Boc-ethylenediamine using NaCNBH₃ or NaH(OAc)₃ inMeOH, THF, or DCE as solvent gives the secondary amine (70). Reaction of(69) with amine (70) followed by deprotection of the Boc group providescompound (71).

Alternatively, compound of formula (74) may be prepared as shown in FIG.16. Reductive amination of an appropriately substituted aniline withtert-butyl N-(2-oxoethyl)carbamate using NaCNBH₃ or NaH(OAc)₃ in MeOH,THF, or DCE as solvent gives the secondary amine (72). Compound (72) canbe purified by removal of the Boc group followed by acid-base extractionand chromatography to give compounds of structure (73) and thenconverted back to compound (72) by treatment with Boc₂O. Reaction withintermediate (69) using DMAP as base followed by deprotection of the Bocgroup with (for example) ethereal HCl and substitution (if required)gives compound (74).

Compound (78) may be prepared as shown in FIG. 17. Compound (75) can beprepared from 7-azaindole according to literature procedures.Introduction of the piperazine can be accomplished by meltingN-benzylpiperidine with intermediate (75) to give intermediate (76).Removal of the benzyl protecting group can be accomplished using (forexample) hydrogenation in the presence of Pd—C in methanol. Coupling ofa Boc-protected amino acid with intermediate (77) can be accomplishedusing (for example) EDCI or PyBrop, followed by deprotection of the Bocgroup to give compound (78).

FIG. 18 shows preparation of compound (81). Compound (79) can beprepared from 2-aminopyridine similar to literature procedure (A. R.Katritzky et al., J. Org. Chem., 2003, 68, 4935-4937). Deprotection ofthe Boc group using (for example) ethereal HCl gives intermediate (80).The piperazine can be coupled to N-protected amino acids using (forexample) EDCI or PyBrop followed by deprotection to give final compound(81).

FIG. 19 illustrates the preparation of 5- and 6-substituted indazole(86). Substituted nitro indazole (82), where R⁵ and R⁷ are substituentswhich are suitable for use in the subsequent reactions, may be reducedto amino indazole (83) using standard conditions (for example catalytichydrogenation, zinc/acetic acid, Fe/HCl, SnCl₂/MeOH or FeSO₄ in water).Amino indazole (83) can react with compound (84) (for example,bis(2-chloroethyl)amine) in the presence of an acid scavenger (forexample, Na₂CO₃, K₂CO₃, or the like) to afford the cyclized product(85). This reaction is performed in a suitable solvent (for example,ethanol) by heating at about 50-150° C. The resulting piperazinecompound (85) is then acylated by a suitable acid (for example aprotected amino acids) which may be introduced using a variety ofstandard peptide coupling procedures under solution phase or solid phaseconditions. The coupling product may require a separate deprotectionstep to remove any protecting groups in R to afford the product (86).For example, a Boc protecting group may be removed by treating with astrong acid such as trifluoroacetic acid (TFA) or hydrochloric acid inthe presence of an inert solvent such as dichloromethane or methanol.Removal of a Cbz group can be carried out by catalytic hydrogenationwith hydrogen in the presence of a palladium catalyst or by transferhydrogenation. An Fmoc group can be removed with a low boiling pointamine (for example piperidine or the like) in a solvent such as DMF.

FIG. 20 describes the synthesis of 3-alkyl and 3-aryl substitutedindazole (90). The iodo intermediate (87) can be prepared by theprocedures shown in FIG. 20. Compound (87) is protected using a suitableprotecting group and treated with an alkyl or aryl boronic acid or esterand a suitable Pd catalyst, for example, Pd(PPh₃)₄, to afford thedesired 3-substituted intermediate (89) which is then deprotected togive compound (90).

The preparation of 3-amino substituted indazoles is outlined in FIG. 21.Compound (92) can be prepared by reacting a suitably monoprotectedpiperazine intermediate with a compound (91), where X is a suitableleaving group (for example, bromo, iodo or OTf), via Pd or Cu-mediatedcoupling (Buchwald et al. (2000), J. Org. Chem., 65, 1144; Hartwig etal. (1998) Angew. Chem. Int. Ed. Eng., 37, 2046) to furnish intermediate(92). Removal of the protecting group followed by amide coupling with anacid affords compound (93), which is then treated with hydrazine to give3-amino indazole intermediate (94). Selective protection of the N-1nitrogen affords compound (95), which can be acylated with an acidhalide in an inert solvent (for example, dichloromethane, or the like)to afford the amide (96). An organic base, such asdiisopropylethylamine, triethylamine, pyridine, or DMAP, may be added asan acid scavenger to facilitate the coupling reaction. Transformation ofcompound (96) into compound (97) can be accomplished by removal of theprotecting groups.

FIG. 22 describes a synthesis of a particular class of compounds bearingan isoquinoline ring. Compound (100) can be prepared by reactingsuitably mono-protected piperazine (99) with an isoquinoline compoundsubstituted with a leaving group X, where X is halide (for example,chloro, bromo, and iodo), or sulfonate (for example OSO₂CF₃), in thepresence of a base and a palladium or a copper catalyst, according toknown methods. Removal of the protecting group Pg of compound (100)affords an amine intermediate, which can be conveniently converted tocompound (101) by amide coupling with acids followed by optional removalof protecting groups as described in FIG. 19.

FIG. 23 presents a synthesis of a particular class of pyrimidinesbearing a substituent at the 5-position. Compound (104) can be preparedby S_(N)Ar reaction between a suitably mono-protected piperazinecompound (103) and a 4-chloro substituted pyrimidine intermediate (102),where X is Br or I, in the presence of an acid scavenger (for example,diisopropylethylamine or triethylamine). Removal of the protecting groupPg followed by amide coupling with an acid (106) affords intermediate(107). Intermediate (107) can react with various coupling components(108) via metal-mediated reactions to furnish product (109). Forexample, compounds bearing an O- or S-linked substituent at the5-position of the pyrimidine ring can be prepared by reactions betweenintermediate (107) and an alcohol or thiol in the presence of a base(for example, Cs₂CO₃) and a Cu catalyst (for example, CuCl, CuI, or thelike) under modified Ullman coupling conditions (Wolter, M. et. al. Org.Lett. 2002, 4, 973-976). In some cases, an additive (for example,2,2,6,6-tetramethyl-heptane-3,5-dione, pentane-2,4-dione,1,10-phenethroline, or the like) is added to accelerate the reaction.Alternatively, if sodium thiolates are available, base is not requiredfor the reaction. The coupling between compound (107) and a thiol mayalso be accomplished by palladium-catalyzed reactions (Kondo, T. et al.Chem. Rev., 2000, 100, 3205-3220; Zheng, N. et al. J. Org. Chem., 1998,63, 9606-9607). Compounds bearing a N-linked substituent at the5-position of the pyrimidine ring can be prepared by a Pd or Cu mediatedcoupling between intermediate (107) and amines (Buchwald et al. (2000),J. Org. Chem., 65, 1144; Hartwig et al. (1998) Angew. Chem. Int. Ed.Eng. 37, 2046). Compounds bearing an alkyl or an aryl substituent at the5-position of the pyrimidine ring can be prepared by Suzuki coupling(Miyaura, N. Suzuki A. (1995), Chem. Rev. 95, 2457; Org. React. (1997),50, 1) between intermediates (107) and (108), wherein Y is a boronicacid or boronic ester, in the presence of a base (for example, Na₂CO₃and Et₃N), a catalytic Pd(0) species (for example, Pd(PPh₃)₄,Pd(PPh₃)₂Cl₂, Pd₂(dba)₃ and Pd(OAc)₂) and a suitable ligand (such asPPh₃ and AsPh₃). Alternatively, 5-alkyl and aryl substituted pyrimidines(109) may also be prepared by Nigeshi or Kumada couplings betweencompounds (107) and (108), wherein Y—R′ is an organo zinc reagent, inthe presence of a Pd (for example, Pd(PPh₃)₄) or Ni (for exampleNi(acac)₂) catalyst. Alternatively, 5-alkyl and aryl substitutedpyrimidines (109) may also be prepared by Stille coupling betweencompounds (107) and (108), wherein Y—R′ is an organostannane reagent, inthe presence of a Pd catalyst.

FIG. 24 describes an alternate synthesis of compounds bearing anO-linked substituent at the 5-position. Compound (III) can be preparedby Cu-catalyzed coupling of intermediate (110) with benzyl alcohol.Removal of the benzyl group by hydrogenation affords5-hydroxylpyrimidine intermediate (112), which can be converted tocompound (115) by deprotection and amide coupling as described in FIG.19. Alkylation of compound (115) with alkyl halides in the presence of abase (for example, K₂CO₃, Cs₂CO₃, or the like) in an inert solvent (forexample, DMF) provides the desired compound (116).

Compound (125) can be synthesized as described in FIG. 25. Anappropriately substituted pyrimidine (119) may be prepared by thecondensation of a corresponding substituted malonic acid diester (117)and a corresponding substituted formamidine (118) in the presence of abase (for example, NaOEt). Treatment of pyrimidine (119) with ahalogenating agent (for example, POCl₃ or POBr₃) affords the dihalide(120). Displacing one of the halogens with protected piperazine (121)gives the mono-substituted compound (122), which can be converted tocompound (123) by reduction (for example, catalytic hydrogenation) ofthe second halogen. Transformation of compound (123) into desiredcompound (125) can be accomplished by the procedures described in FIG.22.

FIG. 26 illustrates an approach to preparation of 5,6-disubstitutedpyrimidines (129). Treatment of compound (126) with a nucleophile (forexample, an amine) either neat or in the presence of a base can give theSNA, product (127). Alternatively, compound (126) can be converted tocompound (127) via various metal mediated coupling reactions such asdescribed in FIG. 23. Transformation of compound (127) into desiredcompound (129) can be accomplished by a sequence of deprotection, amidecoupling and optional deprotection as described in FIG. 22.

The preparation of compounds with an amino group at the 6-position ofthe pyrimidine ring is shown in FIG. 27. Compounds of formula (131) canbe prepared by palladium catalyzed coupling reactions betweenintermediate (130) and an ammonia equivalent (for example, benzophenoneimine). For a review and leading references for arylation of ammoniaequivalents, see Muci, A. R., Buchwald, S. L., Topics in CurrentChemistry, 2002, 219, 131. Removal of the protecting group Pg² incompound (131) furnishes the amino intermediate (132). Transformation of(132) into desired compound (134) can be accomplished by the proceduresdescribed in FIG. 22.

FIG. 28 summarizes the preparation of compounds of the invention bearinga cinnoline ring. Compound (137) can be prepared by a one-pot processfrom 4-hydroxyl cinnoline (135) and protected piperazine (136) through atriflate intermediate (Cacchi, S. et al. Synlett, 1997, 1400).Sequential removal of the protecting group in compound (137) followed byamide coupling and optional deprotection affords compound (139).

FIG. 29 describes a synthesis of a compound containing a diamino group.Protection of the amino group in compound (140) gives a protectedintermediate, which is subjected to Mitsunobu reaction with pthalimideto furnish compound (141). The phthalimide group can be selectivelyremoved with a base (for example, hydrazine and low boiling pointamines). Acylation of compound (142) with a acids using standard peptidecoupling procedures followed by removal of the protecting group affordsthe product (144).

As shown in FIG. 30, the protecting group in compound (141) canalternatively be first selectively removed under known conditions togive compound (146), which can be coupled with an acid to afford theamide (147). Removal of the phthalimide group with a base (for example,hydrazine and a low boiling point amine) leads to the product (148).

FIG. 31 summarizes a preparation of 1-substituted quinolizinones (153).Treatment of compound (149) with an organometallic base (for example,n-BuLi) followed by quenching with 2-ethoxymethylenemalonic acid diethylester yields the Michael addition product (150). Cyclization occurs whenheating compound (150) in an inert solvent (for example, xylene) to giveintermediate (151). The carboxylate group in intermediate (151) may beremoved by heating in an acidic solution (for example, aqueous HCl orH₂SO₄ solution). Sequential removal of the protecting group in compound(152) followed by amide coupling and optional deprotection affordscompound (153).

FIG. 32 describes a synthesis of compounds with a 4-hydroxyl piperidinelinker. Compound (155) can be prepared by S_(N)Ar reaction between asuitably N-protected 4-hydroxyl piperidine compound and a substitutedquinazoline intermediate (154) where X is leaving group (for example Clor Br), in the presence of a base (for example, NaH or triethylamine) ina suitable solvent such as DMF, THF etc. Removal of the protecting groupPg in compound (155) followed by amide coupling with an acid andoptional deprotection affords the desired compound (156).

Synthesis of compounds with a tetrahydropyridine linker is described inFIG. 33. Compound (157), where Pg is an appropriate protecting group, istreated with an organometallic agent (for example, LDA) andN-phenyltrifluoromethanesulfonimide to give the triflate (158)(Eastwood, P. R. (2000), Tetrahedron Lett., 3705). Conversion oftriflate (158) to the corresponding borinate ester (159) is accomplishedby reacting with a suitable diboron species, such asdipinacolatodiboron, or other electrophilic source of boron, with anappropriate palladium catalyst. Ester (159) is then reacted withcompound (160), where X is a leaving group (for example, chloro, bromo,iodo or OTf), under palladium catalysis to give intermediate (161).Alternatively, the cross=coupling reaction also can be performed in thereverse direction by switching the leaving group and boron species. Forexample, intermediate (161) can be prepared by the reaction betweencompound (160), where X is a boronic acid or ester, with the triflate(158). Sequential removal of the protecting group in compound (161)followed by amide coupling and optional deprotection affords compound(162).

FIG. 34 describes the preparation of 5,6-disubstituted pyrrolopyrimidine(175). Compound (168) can be obtained from commercial sources or can beprepared by literature methods (for example, Eger, K. et al. (1987), J.Heterocycl. Chem. 24, 425-430; Roth, H. J. et al. (1975), Arch. Pharm.308, 179-185; Pichler, H. et al. (1986), Liebigs Ann. Chem. 1986,1485-1505). Condensation of compound (168) with formic acid at elevatedtemperature affords intermediate (169) (Traxler, P. M. et al. (1996), J.Med. Chem., 39, 2285-2292). Treating compound (169) with a halogenatingagent (for example, POCl₃) yields the halide (170). Removal of theprotecting group Pg followed by displacement of the halogen withsuitably protected piperazine (172), either neat or in the presence ofan acid scavenger (for example, diisopropylethylamine or triethylamine),leads to intermediate (173). Transformation of intermediate (173) intocompound (175) can be accomplished by the procedures described in FIG.22.

The preparation of 3-substituted pyrazolopyrimidines (182) is describedin FIG. 35. Compound (176) can be obtained from commercial sources orcan be prepared by literature methods (for example, Hamaguchi, M. et al.(1986), Heterocycles. 24, 2111-2115; MaCall, M. A. et al. (1962), J.Org. Chem. 27, 2433-2439). Condensation of compound (176) with hydrazineaffords the cyano intermediate (177), which can be converted to compound(178) by condensing with formic acid at elevated temperature.Alternatively, compound (96) can first be hydrolyzed to afford theprimary amide, which is then condensed with formamide at elevatedtemperature to give the cyclized product (178). Treatment of (178) witha halogenating agent (for example, POCl₃) gives the halide (179).Transformation of (179) into desired compound (182) can be accomplishedby the procedures described in FIG. 22.

An alternate route to the intermediate (179) for the synthesis of3-substituted pyrazolopyrimidines is shown in FIG. 36. Regioselectivedeprotonation of 4,6-dichloropyrimidine at the C-5 position by treatmentwith an organometallic agent (for example, LDA), followed by quenchingwith aldehyde (184) furnishes the hydroxyl intermediate (185) (Radinov,R. et al. (1986), Synthesis, 11, 886-891; Radinov, R. et al. (1991), J.Org. Chem., 56, 4793-4796). Intermediate (185) can be oxidized with anoxidizing agent (for example, CrO₃ or MnO₂) to give ketone (186).Treatment with hydrazine in an inert solvent such as THF or DCM yieldsthe cyclized product (179). Transformation of (179) into desiredcompound (182) can be accomplished by the procedures described in FIG.22.

As shown in FIG. 37, compound (190) may be prepared in two ways. First,the substitution of heterocyclic core (188) with aminoamido piperazine(189) followed by deprotection with acid affords the desired product(190). Second, substitution of the heterocyclic core (188) withN-protected piperazine (191) gives the intermediate (192), which wassubject to deprotection, coupling with amino acid (193) and deprotectionagain with acid to provide the final compound (190). The halide (188)may be obtained from commercial sources or prepared by means known tothose in the art.

The tetrahydropyrado[2,3-d]pyrimidine derivative (199) may be preparedas shown in FIG. 38. A 2-amino-3-pyradocarboxylic acid (194) was heatedwith formamide to give the 4-hydroxypyradopyrimidine derivative (195),which was subject to chlorination with (for example) POCl₃ to afford the4-chloro pyridopyrimidine derivative (196). S_(N)Ar reactions of thecompounds (196) with 1-Boc-piperazine gave the intermediates (197).Reduction of the intermediate (197) in the presence of catalytic amountof (for example) PtO₂ under hydrogen yielded thetetrahydropyrido[2,3-d]pyrimidine derivatives (198). After deprotection,the compounds (198) were subject to amide coupling with N-protectedamino acids and followed by deprotection with acid to offer the product(199).

Dihydropyrrolo[2,3-d]pyrimidine derivatives may be prepared as shown inFIG. 39. 2-Ethoxycarbonyl-succinic acid diethyl ester (200) was heatedwith formamidine to provide (4,6-Dihydroxypyrimidin-5-yl)-acetic acidmethyl ester (201). Halogenation of compound (201) with (for example)POCl₃ gave the dichloropyrimidine derivative (202). Treating compound(142) with base (e.g., KH) in THF and an electrophile R³X affords theintermediate (203). Reduction with an agent such as DIBAL-H gives thealcohol, which was activated with (for example) MsCl in the presence ofbase (e.g., TEA) to provide the mesylate intermediate (204). Treatingthe mesylate (204) with (for example) p-methoxybenzylamine yielded thedihydropyrrolo[2,3-d]pyrimidine derivative (205). S_(N)Ar reaction ofthe compound (205) with 1-Boc-piperazine formed the intermediate (206).Deprotection of the compound (206) with acid (e.g., TFA) gave the freeamine (207). Amide coupling of the free amine (207) with anappropriately substituted/protected amino acid and followed bydeprotection (if necessary) gave the product (208).

Compound (217) of this invention may be prepared as shown in FIG. 40.Thus palladium-catalyzed cross coupling of boronic acid (213) andproperly substituted aryl halide (214) affords the ester intermediate,which is saponified by hydroxide base leading to acid (215). Thecoupling of acid (215) and amine (216) under standard conditions (e.g.,EDCI, HOBt, etc.) gives the N-protected/substituted advancedintermediate (for example Boc, but any suitable protecting group may beused; see, Greene et al., supra. The N-protected/substitutedintermediate (e.g., Boc) is cleaved if necessary to afford product(217).

Compound (223) may be prepared as shown in FIG. 41. Introduction of aleaving group into the appropriately substituted and commerciallyavailable 4,3,0-heterocycle (218) may be accomplished, for example, bytreatment with a halogenating agent (for example POCl₃) to give thechloride (219). Displacement of the leaving group with an appropriatelysubstituted and protected piperazine (220) (for example Boc, but anysuitable protecting group may be used; see, Greene et al., supra)either, for example, neat or in the presence of base, followed byremoval of the piperazine protecting group (using references from theabove book) can give the advanced quinazoline intermediate (221).Substitution of the piperazine secondary amine may then be accomplishedusing a variety of electrophiles and reaction conditions. The piperazinemay be acylated by a suitably N-substituted or protected amino acid(e.g., Boc, etc. (162)) which may be introduced using a variety ofstandard peptide coupling procedures under both solution phase and solidphase conditions to yield compound (223). For representative examples,see Miklos Bodanszky, ‘Principles of Peptide Synthesis,’Springer-Verlarg, 1993, 2nd Ed., and C. Najera, Synlett, 2002, 9,1388-1403. As above (and if protected) the N-protected amino acid unitmay then be deprotected using representative procedures (e.g., acid, fora Boc-group) referenced in Greene et al., supra, and then manipulated asdesired according to procedures appreciated by those skilled in the art.Alternatively (and as in FIG. 1), the amine (221) may be reacted withany other electrophile, including (but not limited to) epoxides, acidhalides, aldehydes, etc., using procedures known to those in the art oforganic synthesis.

Compounds of formula (229) may be prepared as shown in FIG. 42. AnS_(N)Ar reaction of compound (224) with a protected linker (eg.Boc-piperazine) gives the piperazine (225). An organometallic-mediatedreaction may be used to install an activated acetylene group (226) andtreatment with base (for example, KOtBu) gives the pyrrolopyrimidine(228). Deprotection of the piperazine protecting group (with, forexample in the case of a Boc group, acid) and acylation (with forexample a protected amino acid, followed by deprotection if necessary)gives the desired product (229).

Compounds of formula (234) may be prepared as shown in FIG. 43. Thushalogenation (eg. POCl₃) of a suitably substituted nitropyridone, (230),and subsequent treatment with a vinyl Grignard reagent gives theappropriately halogenated pyrrolopyridine (232). Subsequent S_(N)Ardisplacement (or alternatively a transition metal mediated reaction)with a suitably substituted piperazine derivative (233) gives thedesired product (234).

FIG. 44 illustrates the general preparation of compounds of the formula(242). Acylation of an appropriately substituted aminothiophene (235)(using, for example, formic acid and ammonium acetate under heat) andcyclisation using (for example) formamide and ammonium formate at hightemperature gives the appropriate heterocycle. Halogenation, using (forexample) oxalyl chloride then gives the appropriately halogenatedintermediate (237). This intermediate may then be functionalised inmultiple ways. For example, displacement with an appropriatelysubstituted piperazine (using either heat or transition metal mediatedreactions) will give the desired product (242). Alternatively, the coremay be halogenated, using (for example) an organolithium base and ahalogen source (e.g., NCS, Br₂, I₂, etc.) to give compound (238). In thecase of bromination, both non-regioselective and polybromination areobserved, allowing an entry into more fully substituted andfunctionalised derivatives, (239). These may then be subjected to anynumber of anionic or transition metal-mediated reactions (eg. Suzuki,Stille, Negishi, etc.) to provide further functionality (e.g., (240) or(241)). In all cases, subsequent displacement with an appropriatelysubstituted piperazine (along with subsequent functionalisation, ifdesired) gives rise to the desired products (242).

Compounds (247) may be prepared as shown in FIG. 45.4-Chloropyrrolo[2,3-d]pyrimidine (243) is oxidized with an appropriateoxidizing agent (pyridinium tribromide, for example) in an appropriatesolvent (such as t-butanol), and the resulting gem-dibromide is reducedunder appropriate conditions (using Zn/HOAc, for example) in anappropriate solvent (e.g., MeOH) to give the lactam 244. Reaction ofcompound (244) with a monoprotected diamine (245) (using the Bocprotecting group, for example) in an appropriate solvent (such as IPA orNMP) in the presence or absence of base (such as triethylamine),followed by deprotection with mineral acid (HCl, for example) furnishesintermediate (246). Compound (246) is then combined with (for example) aprotected amino acid (using the Boc protecting group, for example) in anappropriate solvent (such as DCM or DMF) with or without base (such astriethylamine or DIEA) and treated with an appropriate coupling agent(such as DCC, HBTU, or EDCI) to furnish compound (247) afterdeprotection with acid (in the case of a Boc-protecting group.)

FIG. 46 shows the general preparation of compounds (256). A suitablysubstituted thiophenecarboxylic acid (248) may be converted to theprotected aminothiophene (249) by means of a rearrangement, using (forexample) diphenylphosphorylazide in the presence of a suitable base andsolvent (e.g., t-BuOH) at elevated temperature. This can be treated witha suitable malonate derivative (eg. 2-ethoxymethylene malonate) at hightemperature to give the pyridothiophene (250). Halogenation using (forexample) POCl₃ in the presence of base (eg. NEt₃) gives the chloride(251) which may then be treated with an appropriate linker (eg.Boc-piperazine) and appropriately deprotected to give the piperazineintermediate (252). Acylation (using, for example, a suitably protectedamino acid under standard conditions) followed by deprotection (ifneeded), gives the desired product (254). Alternatively, in place ofdeprotection, the ester may be saponified using aqueous basic conditions(e.g., LiOH in water and methanol) to give the acid (255) which may thenbe removed by decarboxylation by heating at high temperature in anappropriate solvent (e.g., diphenyl ether.) After any additional andnecessary deprotection, the desired product is attained (256).

The general preparation of compounds (265) is shown in FIG. 47. Henceprotection of an appropriately substituted halopyridone (257) with (forexample) an acetate group under standard conditions) and a subsequenttransition metal mediated introduction of a functionalized actyylene(258) (for example, using PdCl₂(PPh₃)₂ and CuI) gives the intermediateacetylene (260). Treatment with base (e.g., aqueous Na₂CO₃ and heat)effects cyclisation to give the pyridylfuran core (261). Halogenation,via N-oxidation (eg. mcpba oxidation) and treatment with a halogenatingagent (eg. POCl₃) gives the halide (263), which is then displaced by(for example) piperazine to give (264), and then subsequently furtherfunctionalised by (for example) the introduction of a suitably protectedamino acid, followed by deprotection to give the desired product (265).

FIG. 48 illustrates the general preparation of compound (271). Couplingof the uracil derivative (266) with 3-aminopyrazole (267) in thepresence of base gives the pyrazolopyridone (268). Halogenation (forexample, using POCl₃) gives the halide (269), displacement with asuitably substituted linker (e.g., Boc-piperazine using heat) andremoval of the protecting group (e.g., TFA, for a Boc-group), followedby acylation using (for example) a protected amino acid (followed by theappropriate deprotection) gives the desired product (271).

In order to illustrate the invention, the following examples areincluded. However, it is to be understood that these examples do notlimit the invention and are only meant to suggest a method of practicingthe invention. Persons skilled in the art will recognize that thechemical reactions described may be readily adapted to prepare a numberof other analogs of the invention, and alternative methods for preparingthe compounds of this invention are deemed to be within the scope ofthis invention. For example, the synthesis of non-exemplified compoundsaccording to the invention may be successfully performed bymodifications apparent to those skilled in the art, e.g., byappropriately protecting interfering groups, by utilizing other suitablereagents known in the art other than those described, and/or by makingroutine modifications of reaction conditions. Alternatively, otherreactions disclosed herein or known in the art will be recognized ashaving applicability for preparing other compounds of the invention.

BIOLOGICAL EXAMPLES Example of AKT-1 Kinase Assay

The activity of the compounds described in the present invention may bedetermined by the following procedure: This procedure describes a kinaseassay that measures the phosphorylation of a fluorescently-labeledpeptide by full-length human recombinant active AKT-1 by fluorescentpolarization using a commercially available IMAP kit.

The assay materials come from an IMAP AKT Assay Bulk Kit, product#R8059, from Molecular Devices, Sunnyvale, Calif. The kit materialsinclude an IMAP Reaction Buffer (5×): The diluted 1× IMAP ReactionBuffer contains 10 mM Tris-HCl, pH 7.2, 10 mM MgCl₂, 0.1% BSA, 0.05%NaN3. DTT is routinely added to a final concentration of 1 mMimmediately prior to use.

Also included are IMAP Binding Buffer (5×), and IMAP Binding Reagent.The Binding Solution is prepared as a 1:400 dilution of IMAP BindingReagent into 1× IMAP Binding Buffer.

Fluorescein-labeled AKT Substrate (Crosstide), having the sequence(Fl)-GRPRTSSFAEG. A stock solution of 20 μM is made up in 1× IMAPReaction Buffer.

The plates used include a Costar 3657 (382-well made of polypropyleneand having a white, v-bottom) that is used for compound dilution and forpreparing the compound-ATP mixture. The assay plate was the PackardProxyPlate™-384 F.

The AKT-1 used was made from full-length, human recombinant AKT-1 thatis activated with PDK1 and MAP kinase 2.

The assay procedure starts the preparation of stock solutions ofcompounds at 10 mM in DMSO. The stock solutions and the control compoundare serially diluted 1:2 nine times into DMSO (10 μL of compound+10 μLof DMSO) to give 50× dilution series over the desired dosing range.Then, 2.1-μL aliquots of the compounds in DMSO are transferred to aCostar 3657 plate containing 50 μL of 10.4 μM ATP in 1× IMAP ReactionBuffer containing 1 mM DTT. After thorough mixing, 2.5-μL aliquots aretransferred to a ProxyPlate™-384 F plate.

The assay is initiated by the addition of 2.5-μL aliquots of a solutioncontaining 200 nM of fluorescently-labeled peptide substrate and 4 nMAKT-1. The plate is centrifuged for 1 minute at 1000 g and incubated for60 minute at ambient temperature. The reaction is then quenched by theaddition of 15 μL of Binding Solution, centrifuged again and incubatedfor an additional 30 minutes at ambient temperature prior to reading ona Victor 1420 Multilabel HTS Counter configured to measure fluorescencepolarization.

Several compounds of Formula I tested in the assay showed activity forinhibiting AKT protein kinases, including(2R)-2-amino-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one,(2R)-2-amino-3-(2-napthyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one,and(2R)-2-amino-3-(4-chlorophenyl)-1-(4-thieno[3,2,b]pyridin-7-yl-piperazin-1-yl)-propan-1-one.

PREPARATIVE EXAMPLES

The compounds of the present invention may be prepared either as eitherthe racemate or as a single enantiomer (for example, usingenantiomerically pure reagents. If prepared as the racemate, thecorresponding enantiomers may be isolated by separation of the racemicmixture of on a chiral stationary phase column utilizing normal orreverse phase HPLC techniques. Alternatively, a diastereomeric mixturecan be prepared by treatment of the racemic mixture with an appropriatechiral acid (or suitably activated derivative), for example dibenzoyltartrate or the like (see, for example, Kinbara, K., et. al., J. Chem.Soc., Perkin Trans. 2, 1996, 2615; and Tomori, H., et. al., Bull. Chem.Soc. Jpn., 1996, 3581). The diastereomers would then be separated bytraditional techniques (i.e. silica chromatography, crystallization,HPLC, etc) followed by removal of the chiral auxiliary to affordenantiomerically pure material.

The examples below describe the synthesis of some of the compounds ofthe invention. Unless otherwise indicated all temperature s are setforth in degrees Celsius. Reagents were purchased from commercialsuppliers such as Aldrich Chemical Company, Lancaster, TCI or Maybridge,and were used without further purification unless otherwise indicated.

The reactions set forth below were done generally under a positivepressure of nitrogen or argon or with a drying tube (unless otherwisestated) in anhydrous solvents, and the reaction flasks were typicallyfitted with rubber septa for the introduction of substrates and reagentsvia syringe. Glassware was oven dried and/or heat dried.

HPLC retention times (R_(t)) are reported in minutes. Unless statedotherwise, the following HPLC conditions were used to obtain thereported retention times: column: Waters YMC ODS-AQ, 3.0×50 mm; 5-95%gradient MeCN in water (0.01% HFBA, 1% IPA); flow rate: 1.00 mL/min;detected at 220 nm.

¹H-NMR spectra were recorded on a Varian instrument operating at 400MHz. ¹H-NMR spectra were obtained as CDCl₃ solutions (reported in ppm),using chloroform as the reference standard (7.25 ppm). Other NMRsolvents were used as needed. When peak multiplicities are reported, thefollowing abbreviations are used: s (singlet), d (doublet), t (triplet),m (multiplet), br (broadened), dd (doublet of doublets), dt (doublet oftriplets).

Example 1A

Preparation of 4-piperazinylquinazoline Amino Amides

Step 1:

To a solution of 4-chloroquinazoline (2.0 g, 12.2 mmol) (To be,Masanori, et al., Bioorg. Med. Chem. 2003, 11(3), 383) and DIEA (3.2 mL,18.2 mmol) in 40 mL IPA was added Boc-piperazine (1.96 g, 12.81 mmol).The reaction mixture was heated to reflux and stirred for 20 hours,after which it was cooled to room temperature and concentrated by rotaryevaporation. The residue was dissolved in dichloromethane (DCM) andwashed with 1N NaOH. The organic layer was dried (Na₂SO₄), filtered, andconcentrated by rotary evaporation. The resulting oil was dissolved in25 mL dioxane, and 4M HCl/dioxane (46 mL, 182 mmol) was added dropwise.The suspension was sonicated for 2 minutes and stirred 13 hours at roomtemperature, after which the reaction mixture was concentrated todryness by rotary evaporation. The resulting amine HCl salt wasdissolved in 2N NaOH and extracted with DCM. The organic layer was dried(Na₂SO₄), filtered, and concentrated by rotary evaporation. Theresulting oil was purified on silica (9:1:0.02 DCM/MeOH/NH₄OH) to give4-piperazinylquinazoline as a yellow oil (2.5 g, 96%). ¹H NMR (CDCl₃,400 MHz) δ 8.74 (s, 1H), 7.92-7.86 (m, 2H), 7.76-7.70 (m, 1H), 7.48-7.42(m, 1H), 3.75 (t, J=4.9 Hz, 4H), 3.09 (t, J=4.9 Hz, 4H), 1.89 (br s,1H). R_(t) 0.70. MS (ESI+) [M+H]⁺ 215.

Step 2:

To a Jones tube containing PS-CDI (Argonaut, 1.04 mmol/g, 2.2equivalents) suspended in a solution of the 4-piperazinylquinazoline(1.0 equivalent) in CHCl₃ was added a solid Boc-protected amino acid(1.5 equivalents) (see Example 1B). The reaction mixture was shaken for15 hours at room temperature, after which it was vacuum filtered, theresin rinsed with CHCl₃, and the filtrate concentrated by rotaryevaporation. If necessary, the crude coupled product was purified onsilica (DCM/EtOAc or DCM/MeOH). The resulting Boc-amino amide wasdissolved in minimal dioxane, and 4M HCl/dioxane (10 equivalents) wasadded. The suspension was sonicated 5 minutes and stirred at roomtemperature for 12 hours, after which it was concentrated by rotaryevaporation. The solids were dispersed in ether, isolated by filtrationwith nitrogen pressure, and dried under reduced pressure to give thecorresponding 4-piperazinylquinazoline amino amide as the hydrochloridesalt. If necessary, the hydrochloride salts were free-based with 1NNaOH, extracted with DCM, and the combined organic layers were dried(Na₂SO₄), filtered, concentrated by rotary evaporation, and dried underreduced pressure.

Example 1B

The following amino acids were introduced as Boc-protected amino acidsto the 4-piperazinylquinazoline in Example 1, Step 2:

The compounds described in Examples 2-21 were prepared as described inExample 1, Step 2, using 4-piperazinylquinazoline and the appropriateamino acid shown in Example 1B.

Example 2

(2R)-2-Amino-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

R_(t) 2.15. MS (ESI+) [M+H]⁺ 362.

Example 3

(2R)-2-Amino-4-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-one

R_(t) 2.26. MS (ESI+) [M+H]⁺ 376.

Example 4

(2R)-2-Amino-3-(4-methoxphenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

R_(t) 2.15. MS (ESI+) [M+H]⁺ 392.

Example 5

(2R)-2-Amino-1-(4-quinazolin-4-yl-piperazin-1-yl)-3-(2-thienyl)-propan-1-one

R_(t) 2.10. MS (ESI+) [M+H]⁺ 368.

Example 6

(2R)-2-Amino-3-(3-indolyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one.R_(t) 2.24. MS (ESI+) [M+H]⁺ 401. Example 7

(2R)-2-Amino-3-(2-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

R_(t) 2.23. MS (ESI+) [M+H]⁺ 396.

Example 8

(2R)-2-Amino-3-(3-chlorophenyl-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

R_(t) 2.30. MS (ESI+) [M+H]⁺ 396.

Example 9

(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

R_(t) 2.36. MS (ESI+) [M+H]⁺ 396. ¹H NMR (free base; CDCl₃, 400 MHz) δ8.73 (s, 1H), 7.89 (d, J=8.4 Hz, 1H), 7.78 (d, J=8.4 Hz, 1H), 7.77-7.71(m, 1H), 7.49-7.43 (m, 1H), 7.31-7.26 (m, 2H), 7.23-7.19 (m, 2H), 4.37(br s, 1H), 3.92-3.82 (m, 1H), 3.73-3.57 (m, 6H), 3.39-3.30 (m, 1H),3.29-3.19 (m, 1H), 3.12-3.02 (m, 2H).

Example 10

(2R)-2-Amino-3-(4-fluorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one(12)

R_(t) 2.23. MS (ESI+) [M+H]⁺ 380.

Example 11

(2R)-2-Amino-3-(2-naphthyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

R_(t) 2.39. MS (ESI+) [M+H]⁺ 412.

Example 12

(2R)-2-Amino-2-(2-indanyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-ethan-1-one

R_(t) 2.33. MS (ESI+) [M+H]⁺ 388.

Example 13

(±)-2-(4-Fluorophenyl)-2-piperazinyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-ethan-1-one

R_(t) 2.21 minutes. MS (ESI+) [M+H]⁺ 435.

Example 14

(2R)-2-Amino-3-(2,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

R_(t) 2.39. MS (ESI+) [M+H]⁺ 430.

Example 15

(2R)-2-Amino-3-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

R_(t) 2.43. MS (ESI+) [M+H]⁺ 430.

Example 16

(2R)-2-(N-Methylamino)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

R_(t) 2.15. MS (ESI+) [M+H]⁺ 376.

Example 17

Preparation of(2R)-2-(N,N-Dimethylamino)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

To a solution of(2R)-2-(N-Methylamino)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one(free-base, 14 mg, 0.037 mmol) in 1,2-dichloroethane was added asolution of formaldehyde (37% w/w, 50 μL, 0.61 mmol) and then Na(OAc)₃BH(0.12 mmol). The reaction mixture was stirred at room temperature for 3hours, after which saturated NaHCO₃ was added, and the reaction mixturewas stirred 10 minutes. The suspension was extracted with DCM, and thecombined extracts were dried (Na₂SO₄), filtered, and concentrated byrotary evaporation. The residue was filtered through a Fluorosil plugwith ETOAC, and the filtrate was concentrated by rotary evaporation. Theresulting residue was dissolved in ether and excess 4M HCl/dioxane wasadded. The yellow solids were isolated by filtration with nitrogenpressure and dried under reduced pressure to afford the desired product(7 mg, 41%) as the dihydrochloride salt. R_(t) 2.20. MS (ESI+) [M+H]⁺390.

Example 18

(4-Quinazolin-4-yl-piperazin-1-yl)-((3R)-1,2,3,4-tetrahydro-isoquinolin-3-yl)-methanone

R_(t) 2.21. MS (ESI+) [M+H]⁺ 374.

Example 19

Preparation of(2R)-2-(2-Aminoacetamido)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

(2R)-2-Amino-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one(free-base) was acylated with Boc-glycine and deprotected according toStep 2 of Example 1 to furnish(2R)-2-(2-Aminoacetamido)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one.R_(t) 2.23. MS (ESI+) [M+H]⁺ 419.

Example 20

Preparation of(2R)-2-(3-aminopropionamido)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

(2R)-2-Amino-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one(free-base) was acylated with Boc-homoalanine and deprotected accordingto according to Step 2 of Example 1 to furnish(2R)-2-(3-Aminopropionamido)-3-phenyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one.R_(t) 2.24. MS (ESI+) [M+H]⁺ 433.

For the compounds prepared below, the HPLC conditions used to obtain thereported retention times (minutes) were: column: Waters YMC ODS-AQ,4.6×50 mm; 5-95% gradient MeCN in water (0.01% HFBA, 1% IPA); flow rate:2.00 mL/min; detected at 220 nm.

Example 21

Preparation of(2R)-2-[(2-Aminoethyl)amino]-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one(free-base, 41 mg, 0.104 mmol) was stirred at room temperature withBoc-2-aminoacetaldehyde (16 mg, 0.104 mmol) in 0.7 mL methanol for 1hour, after which NaBH₄ (6 mg, 0.160 mmol) was added. The reactionmixture was stirred for 3 hours and then quenched with 1N NaOH. Thereaction mixture was extracted with DCM, and the combined extracts weredried (Na₂SO₄), filtered, and concentrated by rotary evaporation. Thecrude residue was purified on silica (15:1 DCM/MeOH). The resultingBoc-amine intermediate was treated with 1.4 mL 4M HCl/dioxane andstirred at room temperature for 14 hours, after which the reactionmixture was diluted with ether. The solids were isolated by filtrationwith nitrogen pressure and dried under reduced pressure to give thedesired product (29 mg, 51%) as the trihydrochloride salt. R_(t) 1.69.MS (CI+) [M+H]⁺ 439.

Example 22

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-((2R)-2-methyl-4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one

4-((3R)-3-methylpiperazinyl)quinazoline was prepared in a similarfashion to Step 1 of Example 1, which was then acylated withBoc-4-chloro-D-phenylalanine and deprotected according to Step 2 ofExample 1 to furnish(2R)-2-Amino-3-(4-chlorophenyl)-1-((2R)-2-methyl-4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one.R_(t) 1.72. MS (CI+) [M+H]⁺ 410.

Example 23

Preparation of2-Amino-3-(4-chlorophen)-1-[4-(6-phenylquinazolin-4-yl)-piperazin-1-yl]-propan-1-one

Step 1: A solution of 6-bromoquinazolin-4-ol (1.0 g, 4.44 mmol) in POCl₃(10 mL) was stirred and heated at 110° C. in a sealed tube overnight.The solution was cooled to room temperature and poured onto ice (200 g.)The solution was extracted with DCM (300 mL), washed with water (200mL), dried over Na₂SO₄ and concentrated in vacuo to give the impure6-bromo-4-chloroquinazoline as a brown solid that was not purifiedfurther (1.5 g.) MS (APCI+) [M+H]⁺ 243.1.

Step 2: A solution of the crude product from Step 1 (1.5 g),piperazine-1-carboxylic acid tert-butyl ester (2.29 g, 12.3 mmol) andtriethylamine (2.15 mL, 15.4 mmol) in N-methylpyrrolidinone (50 mL) wasstirred and heated at 80° C. for 2 hours. The solution was cooled toroom temperature, diluted with EtOAc (200 mL), washed with water (3×200mL) and dried over Na₂SO₄. The mixture was purified by silica gel columnchromatography (50% EtOAc/hexanes) to give4-(6-bromo-quinazolin-4-yl)-piperazine-1-carboxylic acid tert-butylester as a colorless oil (1.5 g, 3.8 mmol, 85% from Step 1.) MS (APCI+)[M+H]⁺ 394.9 and 392.9. ¹H NMR (CDCl₃, 400 MHz) δ 8.76 (1H, s), 8.02(1H, s), 7.84-7.78 (2H, m), 3.74 (4H, s), 3.66 (4H, s), 1.51 (9H, s.)

Step 3: HCl (1.0 M in Et₂O, 30 mL) was added to a solution of4-(6-bromo-quinazolin-4-yl)-piperazine-1-carboxylic acid tert-butylester (1.50 g, 3.81 mmol) in MeOH (50 mL) and stirred at roomtemperature overnight. The mixture was concentrated in vacuo to give6-bromo-4-piperazin-1-yl-quinazoline as the bis-hydrochloride salt (1.3g, 93%.) MS (APCI+) [M+H]⁺ 295.1.

Step 4: EDCI.HCl (230 mg, 1.2 mmol), HOBt (160 mg, 1.2 mmol) andBoc-D-4-chlorophenylalanine (240 mg, 1.2 mmol) were added to a stirredsolution of 6-bromo-4-piperazin-1-yl-quinazoline bis-hydrochloride (360mg, 0.98 mmol) and triethylamine (0.30 mL, 1.2 mmol) in DMF (8 mL) atroom temperature under nitrogen. Stirred at room temperature overnight.Diluted with EtOAc (100 mL) and washed with water (3×50 mL.) Dried overNa₂SO₄ and concentrated in vacuo. The mixture was purified by silica gelcolumn chromatography (100% EtOAc) to give[2-[4-(6-bromoquinazolin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester as a yellow solid (320 mg, 57%.) MS (APCI+) [M+H]⁺575.9. ¹H NMR (CDCl₃, 400 MHz) δ 8.75 (1H, s), 7.95 (1H, s), 7.85-7.79(2H, m), 7.30-7.27 (2H, m), 7.17 (2H, d, J 7.1 Hz), 5.39 (1H, app. d, J8.2 Hz), 4.86 (1H, app. d, J 7.3 Hz), 3.83-3.60 (6H, m), 3.36-3.26 (2H,m), 3.04-2.96 (2H, m), 1.43 (9H, s.).

Step 5: THF (5 mL) was added to a stirred mixture of Pd₂ dba₃ (8.0 mg,0.0087 mmol) and triphenylarsine (11 mg, 0.035 mmol) at room temperatureunder nitrogen. The yellow solution was stirred at room temperature for2 minutes and then transferred via cannula to a stirred solution of[2-[4-(6-bromoquinazolin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester (50 mg, 0.087 mmol) and phenylboronic acid (21 mg,0.17 mmol) in ethylene glycol dimethyl ether (5 mL) and aqueous sodiumcarbonate (2M, 5 mL) and stirred and heated at 80° C. under nitrogenovernight. The reaction was cooled to room temperature, extracted intoEtOAc (100 mL), washed with water (50 mL), dried over Na₂SO₄ andconcentrated in vacuo. The mixture was purified by silica gel columnchromatography (100% EtOAc) to give{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-phenylquinazolin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester as an oil (30 mg, 60%.) MS (APCI+) [M+H]⁺ 574.0and 572.0.

Step 6: Trifluoroacetic acid (4 mL) was added to a stirred solution of{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-phenyl-quinazolin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester (30 mg) in DCM (10 mL) at room temperature. Thesolution was stirred at room temperature for 4 hours, quenched withaqueous NaOH (1N, 10 mL), diluted with EtOAc (100 mL) and washed withaqueous NaOH (2×50 mL.) The organic phase was dried over Na₂SO₄ andconcentrated in vacuo to give2-amino-3-(4-chlorophenyl)-1-[4-(6-phenyl-quinazolin-4-yl)-piperazin-1-yl]-propan-1-oneas a colorless oil (23 mg.) ¹H NMR (CDCl₃, 400 MHz) δ 8.77 (1H, s),8.02-8.00 (2H, m), 7.98 (1H, s), 7.64 (2H, d, J 7.4 Hz), 7.53 (2H, t, J7.7 Hz), 7.44 (1H, t, J 7.3 Hz), 7.30 (2H, d, J 8.6 Hz), 7.17 (2H, d, J8.6 Hz), 3.98 (1H, t, J 7.3 Hz), 3.87-3.60 (6H, m), 3.42-3.34 (2H, m),2.97 (1H, dd, J 7.4 and 13.3 Hz), 2.84 (1H, dd, J 7.0 and 13.7 Hz.) Theoil was taken up into THF (10 mL) and treated with HCl (1.0M in Et₂O, 10mL) and concentrated in vacuo to give the bis hydrochloride salt of2-amino-3-(4-chlorophenyl)-1-[4-(6-phenyl-quinazolin-4-yl)-piperazin-1-yl]-propan-1-oneas a solid (21 mg.).

Example 24

Preparation of(2R)-2-amino-3-(4-chlorophenyl)-1-(4-thieno[3,2,b]pyridin-7-yl-piperazin-1-yl)-propan-1-onedihydrochloride

Step 1: NaH (60% in mineral oil, 0.24 g) in DMF (15 mL) was addedThieno[3,2-b]pyridin-7-ol (0.756 g) portionwise. The reaction mixturewas warmed at 40° C. and stirred for 30 minutes. After cooling,N-phenyltrifluoromethanesulfonimide (2.1 g) was added, the reactionmixture was stirred at room temperature for 1 hour, and theBoc-piperazine (1.9 g) was added. The mixture was stirred at 80° C. for2 hours. Ethyl acetate (100 mL) was added and the resulting solution waswashed with brine (2×50 mL), dried over sodium sulfate, concentratedunder reduced pressure and purified by chromatography (1:4 hexane/EtOAc)to give the product as yellow oil 1.32 g (82.5%). R_(t) 2.10 minutes. MS(ESI+) [M+H]⁺ 320.

Step 2: The 4-thieno[3,2-b]pyridin-7-piperazine-1-carboxylic tert-butylester (1.32 g) in DCM (20 mL) was added the 4 N HCl in dioxane (21 mL).The reaction was stirred at room temperature for 10 hours. The solventwas removed under reduced pressure and the resulting amine HCl salt wasdissolved in saturated sodium bicarbonate (20 mL) and extracted with DCM(30 mL). The organic layer was dried (Na₂SO₄), filtered, andconcentrated by rotary evaporation. to give7-piperazin-1-yl-thieno[3,2-b]pyridine as an off-white solid (0.85 g,93.8%). R_(t) 1.40 minutes. MS (ESI+) [M+H]⁺ 220.

Step 3: DIEA (0.07 mL) and HBTU (0.12 g) was added to the solution ofthe (2R)-2-tert-butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid(0.092 g) in THF (5 mL) at 0° C. The mixture was stirred at roomtemperature for 20 minutes, then 7-piperazin-1-yl-thieno[3,2-b]pyridine(0.056 g) was added. The reaction was stirred at room temperature for 1hr. 20 mL of EtOAc was added and the organic layer was separated. Theaqueous layer extracted with EtOAc (20 mL). The combined organic layerwas washed with saturated sodium bicarbonate (20 mL) and dried oversodium sulfate. After removal of solvent, the residue was purified byflash chromatography (EtOAc) to give the product as white foam solid(0.126 g, 98.5%). R_(t) 2.43 minutes. MS (ESI+) [M+H]⁺ 501.

Step 4: The resulting Boc-amino amide (0.056 mg) was dissolved indioxane, and 4M HCl/dioxane (0.5 mL) was added. The suspension wasstirred at room temperature for 3 hours, after which it was concentratedto give the corresponding amino amide as the hydrochloride salt (0.53 g,98%). R_(t) 1.77 minutes. MS (ESI+) [M+H]⁺ 401.

Example 25

Preparation of2-Amino-3-(4-chlorophenyl)-1-(4-thieno[3,2-d]pyrimidin-4-yl-piperazin-1-yl)-propan-1-onedihydrochloride

Step 1: To a solution of Boc-D-Phe(4-Cl)—OH (3.65 g, 12.2 mmol),piperazine (10 g, 116 mmol) in DCM (200 mL) were added HOBT (3.3 g, 24mmol) and EDCI (4.7 g, 25 mmol). The mixture was stirred at roomtemperature for 12 hours. The solution was washed with water, brine anddried over magnesium sulfate. After filtration, the organic solvent wasevaporated and the residue was subject to silica gel chromatography toafford the product[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acidtert-butyl ester (0.58 g, 13%). ¹H NMR (CDCl₃, 400 Hz) δ 7.25 (d, J=8.4Hz, 2H), 7.12 (d, J=8.0 Hz, 2H), 5.52 (d, J=8.4 Hz, 1H), 4.83-4.77 (m,1H), 3.63-3.45 (m, 2H), 3.37-3.32 (m, 1H), 3.08-3.04 (m, 1H), 2.99-2.90(m, 2H), 2.81-2.70 (m, 3H), 2.42-2.38 (m, 1H), 1.41 (s, 9H). MS (ESI+)[M+H]⁺ 369.

Step 2: To a solution of 3-aminothiophene-2-carboxylic methyl ester (20g, 127 mmol) in formic acid (100 mL) was added ammonium acetate (13 g,169 mmol). The mixture was refluxed for 3 hours. After cooling to roomtemperature, the precipitate was filtered, washed with water and driedunder vacuum to afford 3-formylaminothiophene-2-carboxylic acid methylester (20.5 g, 87%). ¹H NMR (DMSO 400 Hz) δ 10.38 (s, 1H), 8.42 (s, 1H),8.00 (d, J=5.2 Hz, 1H), 7.90 (d, J=5.6 Hz, 1H), 3.84 (s, 3H). MS (ESI+)[M+H]⁺ 186.

Step 3: To a mixture of 3-formylaminothiophene-2-carboxylic acid methylester (20.5 g, 111 mmol) and ammonium formate (21 g, 333 mmol) was addedformamide (29.8 g, 662 mmol). The slurry was heated to 140° C. for 10hours. After cooling, the solid was filtered, washed with water anddried under vacuum to afford the product 3H-Thieno[3,2-d]pyrimidin-4-one(12.5 g, 74%). ¹H NMR (DMSO, 400 Hz) δ 12.31 (br, 1H), 8.18 (d, J=5.2Hz, 1H), 8.16 (s, 1H), 7.41 (d, J=5.2 Hz, 1H). MS (ESI+) [M+H]⁺ 153

Step 4: To a solution of DMF (13.2 mL, 170 mmol) in DCM (100 mL) at 0°C. was added oxalyl chloride (22 mL, 252 mmol) in DCM (100 mL) veryslowly over 1 hour. To the resulting white gel solution was added the3H-thieno[3,2-d]pyrimidin-4-one (12 g, 79 mmol). The mixture wasrefluxed for 4 hours. After cooling, the mixture was purred into water(500 mL) and extracted with DCM (3×250 mL). Then the organic phase wasdried over magnesium sulfate, filtered and concentrated to afford7-chloro-thieno[3,2-b]pyridine as white solid (13.4 g, 99%). ¹H NMR(CDCl₃, 400 Hz) δ 9.00 (s, 1H), 8.06 (d, J=5.6 Hz, 1H), 7.61 (d, J=5.6Hz, 1H). MS (ESI+) [M+H]⁺ 170

Step 5: The solution of[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acidtert-butyl ester (60 mg, 0.163 mmol) and 4-Chlorothieno[3,2-d]pyrimidine950 mg, 0.293 mmol) in Toluene (5 mL)/TEA (1 mL) was refluxed for 12hours. After the solvent was removed, the residue was subject tochromatography on silica gel to afford[1-(4-chlorobenzyl)-2-oxo-2-(4-thieno[3,2-d]pyrimidin-4-yl-piperazin-1-yl)-ethyl]-carbamicacid tert-butyl ester (71 mg, 86.7%). ¹H NMR (CDCl₃, 400 Hz) δ 8.60 (s,1H), 7.76 (d, J=5.2 Hz, 1H), 7.45 (d, J=5.2 Hz, 1H), 7.26 (d, J=8.4 Hz,1H), 7.15 (d, J=8.4 Hz, 1H), 5.39 (d, J=8.8 Hz, 1H), 4.88-4.82 (m, 1H),3.94-3.76 (m, 4H), 3.69-3.58 (m, 3H), 3.25-3.22 (m, 1H), 2.98 (d, J=7.2Hz, 2H), 1.43 (s, 9H). MS (ESI+) [M+H]⁺ 503.

Step 6: To a solution of[1-(4-chlorobenzyl)-2-oxo-2-(4-thieno[3,2-d]pyrimidin-4-yl-piperazin-1-yl)-ethyl]-carbamicacid tert-butyl ester in DCM (2 mL) was added HCl in Dioxane (4M, 1 mL).The mixture was stirred at room temperature for 4 hours. The solvent wasremoved to afford2-amino-3-(4-chlorophenyl)-1-(4-thieno[3,2-d]pyrimidin-4-yl-piperazin-1-yl)-propan-1-onedihydrochloride quantitatively. MS (ESI+) [M+H]⁺ 402.

Example 26

Preparation of2-amino-3-(4-chlorophenyl)-1-[4-(5H-pyrrolo[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a solution of 4,6-dichloro-5-aminopyrimidine (1 g, 6.1 mmol)in TEA (2 mL) and toluene (10 mL) was added 1-Boc-piperazine (2.3 g,12.3 mmol). The mixture was refluxed for 12 hours. The solvent wasremoved and the residue was subject to chromatography on silica gel toafford the product4-(5-amino-6-chloropyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester (1.9 g, 99%). ¹H NMR (CDCl₃, 400 Hz) δ 8.16 (s, 1H),3.87 (s, 2H), 3.56 (m, 4H), 3.29 (m, 4H), 1.49 (s, 9H). MS (ESI+) [M+H]⁺314.

Step 2: To a solution of4-(5-amino-6-chloropyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester (1 g, 3.19 mmol) and TMS-acetylene (1.5 g, 15 mmol) inTEA (10 mL) and THF (30 mL) were added PdCl₂(PPh₃)₂ (0.33 g, 0.47 mmol)and CuI (0.1 g, 0.53 mmol) under N₂. The mixture was heated to 80° C.for 20 hours. The solvent was removed and the residue was subject tochromatography on silica gel to afford the product4-(5-Amino-6-trimethylsilanylethynyl-pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (0.3 g, 25%). ¹H NMR (CDCl₃, 400 Hz) δ 8.00 (s,1H), 3.74 (s, 2H), 3.28-3.25 (m, 4H), 3.02-2.99 (m, 4H), 1.20 (s, 9H),0.00 (s, 9H). MS (ESI+) [M+H]⁺ 376.

Step 3: To a solution of tBuOK (0.063 g, 0.56 mmol) in NMP (4 mL) wasadded4-(5-Amino-6-trimethylsilanylethynyl-pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (0.1 g, 0.27 mmol) in NMP (1 mL) under N₂. Themixture was vigorously stirred at room temperature for 4 hours. Thereaction was quenched with water (1 mL) and ethyl acetate (50 mL). Theorganic phase was washed brine and water until NMP was gone, then driedover MgSO₄, filtered and concentrated. The residue was subject tochromatography on silica gel to afford the product4-(5H-Pyrrolo[3,2-d]pyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester (43 mg, 53%). ¹H NMR (CDCl₃, 400 Hz) δ 9.47 (s, 1H),8.49 (s, 1H), 7.37 (d, J=2.4 Hz, 1H), 6.62 (d, J=2.8 Hz, 1H), 3.85-3.82(m, 4H), 3.61-3.58 (m, 4H). 1.49 (s, 9H). MS (ESI+) [M+H]⁺ 304.

Step 4: To a solution of4-(5H-Pyrrolo[3,2-d]pyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester (43 mg, 0.14 mmol) in DCM (4 mL) was added HCl inDioxane (4M, 1 mL). The mixture was stirred at room temperature for 4hours. The solvent was removed and the residue was treated with TEA (2mL), Boc-D-Phe(4-Cl)—OH (43 mg, 0.14 mmol), HOBT (30 mg, 0.222 mmol) andEDCI (41 mg, 0.214 mmol) in DCM (5 mL). The mixture was stirred at roomtemperature for 12 hours. The solvent was removed and the residue wassubject to chromatography on silica gel to give product{1-(4-chlorobenzyl)-2-oxo-2-[4-(5H-pyrrolo[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester (30 mg, 44%). ¹H NMR (CDCl₃, 400 Hz) δ 9.85 (s,1H), 8.48 (s, 1H), 7.37 (s, 1H), 7.25-7.23 (d, J=8.4 Hz, 2H), 7.13-7.11(d, J=8.4 Hz, 2H), 6.60 (s, 1H), 5.39-5.37 (d, J=8.8 Hz, 1H), 4.83-4.78(m, 1H), 3.81-3.43 (m, 7H), 3.31-3.27 (m, 1H), 3.00-2.91 (m, 2H), 1.41(s, 9H). MS (ESI+) [M+H]⁺ 485.

Step 5: To a solution of{1-(4-chlorobenzyl)-2-oxo-2-[4-(5H-pyrrolo[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester in DCM (4 mL) was added HCl in Dioxane (4M, 1 mL)and stirred for 4 hours. The solvent was removed to afford2-Amino-3-(4-chlorophenyl)-1-[4-(5H-pyrrolo[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride quantitatively. MS (ESI+) [M+H]⁺ 385.

Example 27

Preparation of2-Amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[3,2-b]pyridin-7-yl)-piperazin-1-yl]-propan-1-one

Step 1: To a solution of 4-hydroxy-3-nitro-pyridine (2 g, 14 mmol) inPOCl₃ (6 mL) was added PCl₅ (2.5 g, 12 mmol). The mixture was heated toreflux for 3 hours. The solvent was evaporated and the residue wascooled with ice-water and vigorously stirred with water (3 mL) and CHCl₃(6 mL). The aqueous was extracted CHCl₃ (5×5 mL). The organic phase wascombined and dried over MgSO₄. After filtration, the solvent was removedto afford the product 4-Chloro-3-nitro-pyridine (2.24 g, 99%). ¹H NMR(CDCl₃, 400 Hz) δ 9.13 (s, 1H), 8.70 (d, J=5.2 Hz, 1H), 7.55 (d, J=5.6Hz, 1H). MS (ESI+) [M+H]⁺ 159.

Step 2: To a solution of 4-chloro-3-nitro-pyridine (2 g, 13 mmol) in dryTHF (100 mL) under N₂ at −78° C. was added excess vinyl magnesiumbromide (1.0M, 40 mL, 40 mmol). The mixture was stirred at −20° C. for 8hours before the reaction was quenched with 20% NH₄Cl (75 mL). Theaqueous phase was extracted with ethyl acetate (3×100 mL). The combinedorganic layer was dried over MgSO₄, filtered and concentrated. Theresidue was subject to chromatography on silica gel to afford7-Chloro-1H-pyrrolo[3,2-b]pyridine (0.3 g, 16%). ¹H NMR (CD₃OD, 400 Hz)δ 8.22 (d, J=5.2 Hz, 1H), 7.64 (d, J=3.2 Hz, 1H), 7.23 (d, J=5.2 Hz,1H), 6.67 (d, J=3.2 Hz, 1H). MS (ESI+) [M+H]⁺ 153.

Step 3: To a solution of 7-Chloro-1H-pyrrolo[3,2-b]pyridine (40 mg,0.262 mmol) in xylene (4 mL) and TEA (1 mL) was added[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acidtert-butyl ester (0.1 g, 0.27 mol). The mixture was refluxed for 6 days.The solvent was removed under vacuum and the residue was subject topurification by HPLC to afford{1-(4-chlorobenzyl)-2-oxo-2-[4-(1H-pyrrolo[3,2-b]pyridin-7-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester (10 mg, 8%). ¹H NMR (CDCl₃, 400 Hz) δ 11.81 (s,1H), 7.95 (d, J=6.4 Hz, 1H), 7.40 (s, 1H), 7.15 (d, J=8.00 Hz, 2H), 6.46(d, J=6.8 Hz, 2H), 5.42 (d, J=8.4 Hz, 1H), 4.82 (d, J=7.2 Hz, 1H),3.87-3.28 (m, 8H), 2.98 (d, J=6.4 Hz, 2H), 1.41 (s, 9H). MS (ESI+)[M+H]⁺ 484.

Step 4: To a solution of{1-(4-chlorobenzyl)-2-oxo-2-[4-(1H-pyrrolo[3,2-b]pyridin-7-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester in DCM (4 mL) was added HCl in Dioxane (4M, 1 mL).The mixture was stirred at room temperature for 4 hours. The solventremoved to afford the product2-Amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[3,2-b]pyridin-7-yl)-piperazin-1-yl]-propan-1-onequantitatively. MS (ESI+) [M+H]⁺ 384.

Example 28

Preparation of2-Amino-3-(4-chlorophenyl)-1-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one

Step 1: To a solution of LDA (1.8M, 20.6 mL, 37.1 mmol) in THF (65 mL)at −78° C. was added 4-chloro-thieno[3,2-d]pyrimidine (5.26 g, 31 mmol)in THF (50 mL) dropwise over 1 hour. After stirring for 20 minutes, I₂(12.7 g, 50 mmol) in THF (40 mL) was added to the mixture at −78° C.dropwise. The mixture was stirred at the same temperature for 20 minutesand then warmed up to room temperature for 2 hours. The mixture waspoured into water (100 mL) and stirred for 30 minutes. The solid wasfiltered and washed with water and Hexane-Hexanes/DCM (50:1) to affordthe product 4-Chloro-6-iodothieno[3,2-d]pyrimidine (6.86 g, 75%). ¹H NMR(DMSO, 400 Hz) δ 8.97 (s, 1H), 8.15 (s, 1H).

Step 2: To a solution of 4-chloro-6-iodothieno[3,2-d]pyrimidine (0.22 g,0.742 mmol) in DCE (5 mL)/TEA (2 mL) was added[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acidtert-butyl ester (25 mg, 0.68 mmol). The mixture was refluxed for 2hours. The solvent was removed and the residue was subject tochromatography on silica gel to afford the product{1-(4-chlorobenzyl)-2-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (44 mg, 95%). ¹H NMR (CDCl₃, 400 Hz) δ 8.47 (s,1H), 7.64 (s, 1H), 7.27-7.25 (d, J=8.4 Hz, 2H), 7.17-7.15 (d, J=8.0 Hz,2H), 5.44 (d, J=8.8 Hz, 1H), 4.85-4.81 (m, 1H), 3.86-3.50 (m, 7H),3.24-3.19 (m, 1H), 2.99-2.97 (d, J=7.2 Hz, 2H), 1.43 (s, 9H). MS (ESI+)[M+H]⁺ 628.

Step 3: To a solution of{1-(4-chlorobenzyl)-2-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester in DCM (4 mL) was added HCl in Dioxane (4M, 1 mL).The mixture was stirred at room temperature for 4 hours. The solvent wasremoved to afford the product2-Amino-3-(4-chlorophenyl)-1-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onequantitatively. MS (ESI+) [M+H]⁺ 528.

Example 29

Preparation of2-Amino-3-(4-chlorophenyl)-1-[4-(6-prop-1-ynyl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a solution of ZnBr₂ (70 mg, 0.311 mmol) in THF (2 mL) wasadded propargyl magnesium bromide (0.5M, 0.6 mL, 0.3 mmol) at roomtemperature. After stirring for 20 minutes, the6-iodothieno[3,2-d]pyrimidine (50 mg, 0.08 mmol) was added. The mixturewas flushed with N₂ and PdCl₂(dppf) was added. The mixture was stirredat room temperature under N₂ for 12 hours. After filtration, thefiltrate was concentrated and the residue was subject to chromatographyon silica gel to afford the product{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-prop-1-ynyl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester (17 mg, 40%). ¹H NMR (CDCl₃, 400 Hz) δ 8.56 (s,1H), 7.37 (s, 1H), 7.27-7.25 (d, J=8 Hz, 2H), 7.16-7.14 (d, J=8.4 Hz,2H), 5.37-5.35 (d, J=8.4 Hz, 1H), 4.85-4.82 (m, 1H), 3.86-3.51 (m, 7H,3.23-3.20 (m, 1H), 3.00-2.94 (m, 2H), 2.15 (s, 3H), 1.42 (s, 9H). MS(ESI+) [M+H]⁺ 540.

Step 2: To a solution of{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-prop-1-ynyl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester in DCM (4 mL) was added HCl in Dioxane (4M, 1 mL).The mixture was stirred for 4 hours. The solvent was removed to affordthe product2-Amino-3-(4-chlorophenyl)-1-[4-(6-prop-1-ynyl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride quantitatively. MS (ESI+) [M+H]⁺ 440.

Example 30

Preparation of2-Amino-3-(4-chlorophenyl)-1-[4-(6-thiophen-3-yl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a solution of{1-(4-chlorobenzyl)-2-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (50 mg, 0.080 mmol) in DMF (3 mL) were added 2MNa₂CO₃ (0.1 mL) and 3-thiophenyl boronic acid (15 mg, 0.117 mmol). Themixture was bubbled N₂ for 20 minutes and then Pd(PPh₃)₄ (10 mg, 0.012mmol) was added. The mixture was heated to 90° C. for 12 hours. Thesolvent was removed under vacuum and the residue was subject tochromatography on silica gel to afford the product{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-thiophen-3-yl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester (16 mg, 34.4%). ¹H NMR (CDCl₃, 400 Hz) δ 8.57 (s,1H), 7.70-7.42 (m, 4H), 7.27 (d, J=8.4, 2H), 7.16 (d, J=8.4 Hz, 2H),5.36 (d, J=8.8 Hz, 1H), 4.87-4.84 (m, 1H), 3.91-3.59 (m, 7H), 3.27-3.24(d, J=11.2 Hz, 1H), 3.00-2.99 (d, J=7.2 Hz, 2H), 1.43 (s, 9H). MS (ESI+)[M+H]⁺ 584.

Step 2: To a solution of product{1-(4-chlorobenzyl)-2-oxo-2-[4-(6-thiophen-3-yl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester in DCM (4 mE) was added HCl in Dioxane (4M, 1 mL).The mixture was stirred at room temperature for 4 hours. The solvent wasremoved to afford the product2-Amino-3-(4-chlorophenyl)-1-[4-(6-thiophen-3-yl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride quantitatively. MS (ESI+) [M+H]⁺ 484.

Example 31

Preparation of2-Amino-3-(4-chlorophenyl)-1-[4-(6-methylsulfanyl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a solution of{1-(4-chlorobenzyl)-2-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (50 mg, 0.080 mmol), sodium methylthiolate (12 mg,2.15 mmol) and 1,3-di-tert-butyl-propane-dione 940 mg, 0.22 mmol) waspurged with N₂. NMP (2 mL) and CuCl (5 mg, 0.05 mmol) were added. Themixture was heated to 130° C. for 3 hours. After cooling down, thereaction was diluted with ethyl acetate and filtered. The filtrate waswashed with water, brine and dried over MgSO₄. After filtration, thesolvent was removed and the residue was subject to chromatography onsilica gel to afford the product{1-(4-chlorobenzyl)-2-[4-(6-methylsulfanyl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. (20 mg, 46%). ¹H NMR (CDCl₃, 400 Hz) δ 8.61 (s,1H), 7.57 (s, 1H), 7.28-7.26 (d, J=6.4 Hz, 2H), 7.18-7.16 (d, J=8.4 Hz,2H), 5.33-5.31 (d, J=8.8 Hz, 1H), 4.81-4.79 (m, 1H), 4.01-3.71 (m, 7H),3.24-3.19 (m, 1H), 3.03-2.98 (m, 2H), 2.74 (s, 3H), 1.43 (s, 9H). MS(ESI+) [M+H]⁺ 548.

Step 2: To a solution of{1-(4-chlorobenzyl)-2-[4-(6-methylsulfanyl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester in DCM (4 mL) was added HCl in Dioxane (4M, 1 mL).The mixture was stirred at room temperature for 4 hours. The solvent wasremoved to afford the product2-Amino-3-(4-chlorophenyl)-1-[4-(6-methylsulfanyl-thieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride quantitatively. MS (ESI+) [M+H]⁺ 448.

Example 32

Preparation of4-{4-[2-Amino-3-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-thieno[3,2-d]pyrimidine-6-carbonitriledihydrochloride

Step 1: To a solution of{1-(4-chlorobenzyl)-2-[4-(6-iodothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (50 mg, 0.080 mmol) in Pyridine (5 mL) was addedCuCN (20 mg, 0.223 mmol). The mixture was refluxed under N₂ for 12hours. The solvent was removed and the residue was subject tochromatography on silica gel to afford{1-(4-chlorobenzyl)-2-[4-(6-cyanothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (29 mg, 69%). ¹H NMR (CDCl₃, 400 Hz) δ 8.65 (s,1H), 7.95 (s, 1H), 7.28-7.26 (d, J=7.6 Hz, 2H), 7.18-7.16 (d, J=8.4 Hz,2H). 5.35-5.33 (d, J=8.4 Hz, 1H), 4.85-4.80 (m, 1H), 3.91-3.56 (m, 7H,3.24-3.19 (m, 1H), 3.01-2.99 (d, J=8.0 Hz, 2H), 1.44 (s, 9H). MS (ESI+)[M+H]⁺ 527.

Step 2: To a solution of{1-(4-chlorobenzyl)-2-[4-(6-cyanothieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester in DCM (4 mL) was added HCl in Dioxane (4M, 1 mL).The mixture was stirred at room temperature for 10 hours. The solventwas removed and the residue was subject to purification by HPLC toafford the product4-{4-[2-Amino-3-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-thieno[3,2-d]pyrimidine-6-carbonitriledihydrochloride (12 mg, 44%). MS (ESI+) [M+H]⁺ 427.

Example 33

Preparation of2-Amino-3-(4-chlorophenyl)-1-[4-(6-methylthieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a solution of 4-chloro-6-iodothieno[3,2-d]pyrimidine (0.5 g,1.7 mmol) in DCE (5 mL)/TEA (1 mL) was added 1-Benzyl-piperazine (0.3 g,1.69 mmol). The mixture was refluxed for 1 hour. The solvent was removedand the residue was subject to chromatography on silica gel to afford(4-Benzyl-piperazin-1-yl)-6-iodothieno[3,2-d]pyrimidine (0.65 g, 88%).¹H NMR (CDCl₃, 400 Hz) δ 8.47 (s, 1H) 7.60 (s, 1H), 7.35-7.23 (m, 5H),3.95-3.92 (m, 4H), 3.51 (s, 2H), 2.59-2.54 (m, 4H). MS (ESI+) [M+H]⁺437.

Step 2: To a suspension of ZnBr₂ (0.5 g, 2.2 mmol) dried under vacuum inTHF (10 mL) was added MeMgBr (3M, 0.6 mL, 1.8 mmol) at room temperaturedropwise. After addition, the mixture was stirred for 1 hour, then(4-Benzyl-piperazin-1-yl)-6-iodothieno[3,2-d]pyrimidine (0.4 g, 0.92mmol) was added followed by PdCl₂(dppf) (30 mg) under N₂. The mixturewas heated to 60° C. for 2 hours. The reaction was quenched with water.The organic phase was separated and dried over MgSO₄. After filtration,the solvent was removed and the residue was subject to chromatography onsilica gel to give the product4-(4-Benzyl-piperazin-1-yl)-6-methylthieno[3,2-d]pyrimidine (0.16 g,54%). ¹H NMR (CDCl₃, 400 Hz) δ 8.52 (s, 1H), 7.35-7.26 (m, 5H), 7.07 (s,1H), 3.98-3.95 (m, 4H), 3.56 (s, 2H), 2.59-2.57 (m, 7H). MS (ESI+)[M+H]⁺ 325.

Step 3: To a solution of4-(4-Benzyl-piperazin-1-yl)-6-methylthieno[3,2-d]pyrimidine (65 mg, 0.20mmol) in MeOH (10 mL) was added Pd/C (10%, 20 mg) and two drop of TFA.The mixture was stirred under H₂ balloon for 4 hours. The catalyst wasfiltered off and the filtrate was concentrated. The residue wasdissolved in DCM (6 mL) and TEA (2 mL), then Boc-D-Phe(4-Cl)—OH (59 mg,0.20 mmol) was added, followed by HOBT (50 mg, 0.37 mmol) and EDCI (74mg, 0.39 mmol). The mixture was stirred at room temperature for 12hours. The solvent was removed and the residue was subject tochromatography on silica gel to give the product{1-(4-chlorobenzyl)-2-[4-(6-methylthieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (60 mg, 59%). ¹H NMR (CDCl₃, 400 Hz) δ 8.54 (s,1H), 7.28-7.26 (d, J=7.2 Hz, 2H), 7.17-7.15 (d, J=8.4 Hz, 2H), 7.08 (s,1H), 5.37-5.35 9d, J=8.8 Hz, 1H), 4.86-4.84 (m, 1H), 3.87-3.57 (m, 7H),3.24-3.22 (m, 1H), 3.00-2.98 (d, J=7.2 Hz, 2H), 2.63 (s, 3H), 1.43 (s,9H). MS (ESI+) [M+H]⁺ 516.

Step 4: To a solution of{1-(4-chlorobenzyl)-2-[4-(6-methylthieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester in DCM (4 mL) was added HCl in Dioxane (4M, 1 mL).The mixture was stirred for 4 hours. The solvent was removed to affordthe product2-Amino-3-(4-chlorophenyl)-1-[4-(6-methylthieno[3,2-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride quantitatively. MS (ESI+) [M+H]⁺ 416.

Example 34

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-5-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: A mixture of 5-aminoindazole (2.53 g, 19.0 mmol),bis(2-chloroethyl)amine hydrochloride (3.60 g, 20.1 mmol) and ethanol(30 mL) was heated at reflux overnight. The mixture was allowed to coolto room temperature. Na₂CO₃ (2.14 g, 20.2 mmol) was added and thereaction mixture heated at reflux for 8 hours. After cooling, themixture was filtered and the filtrate evaporated in vacuo. The residuewas dissolved in 1 N HCl (100 mL) and extracted with DCM (2×50 mL). Theaqueous phase was made basic with 4 N NaOH (30 mL) and extracted withEtOAc (2×100 mL). The combined organic layers were washed with brine,dried and concentrated. The residue was purified by columnchromatography (20:1 DCM/MeOH to 20:1:0.5 DCM/MeOH/Et₃N) to5-piperazin-1-yl-1H-indazole (1.26 g, 33%) as a brown solid. ¹H NMR(DMSO-d₆, 400 MHz) δ 12.80 (s, 1H), 7.89 (s, 1H), 7.40 (d, J=8.8 Hz,1H), 7.16 (dd, J=8.8 Hz, J=2.0 Hz, 1H), 7.07 (s, 1H), 3.17 (s, 1H), 2.99(m, 4H), 2.89 (m, 4H). LCMS (APCI+) m/z 203 [M+H]⁺; Rt=1.33 minutes.

Step 2: To a solution of (D)-Boc-4-chlorophenylalanine (0.119 g, 0.396mmol) and 5-piperazin-1-yl-1H-indazole (0.100 g, 0.494 mmol) in DMF (5mL) was added EDCI (0.152 g, 0.791 mmol), HOBt (0.121 g, 0.791 mmol) andtriethylamine (0.110 mL, 0.791 mmol). The reaction mixture was stirredat room temperature overnight. The mixture was partitioned between waterand EtOAc. The aqueous phase was extracted with EtOAc. The combinedorganic layers were washed with aqueous NaHCO₃, brine, dried andconcentrated. The residue was purified by column chromatography (80:1 to50:1 DCM/MeOH) to give(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (0.176 g, 92%) as a white solid. ¹H NMR (CDCl₃,400 MHz) δ 10.12 (s, 1H), 7.98 (s, 1H), 7.41 (d, J=8.8 Hz, 1H), 7.28 (d,J=8.4 Hz, 2H), 7.16 (d, J=8.4 Hz, 2H), 7.12 (dd, J=8.8 Hz, J=2.0 Hz,1H), 7.07 (s, 1H), 5.46 (m, 1H), 4.88 (m, 1H), 3.74 (m, 2H), 3.53 (m,1H), 3.31 (m, 1H), 3.07 (m, 1H), 2.99 (d, J=6.8 Hz, 2H), 2.91 (m, 2H),2.49 (m, 1H), 1.43 (s, 9H). LCMS (APCI+) m/z 484, 486 [M+H]⁺; Rt=3.01minutes.

Step 3: To a solution of(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (0.176 g, 0.364 mmol) in DCM (10 mL) was added 4 NHCl in dioxane (1 mL). The mixture was stirred at room temperatureovernight and then evaporated. The resulting solid was suspended inisopropyl alcohol-ether (1:5) and stirred for 30 minutes. The mixturewas filtered to give(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-5-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (0.151 g, 91%) as a white solid. ¹H NMR (CD₃OD, 400 MHz)δ 8.24 (s, 1H), 8.03 (s, 1H), 7.75 (d, J=9.2 Hz, 1H), 7.65 (d, J=9.2 Hz,1H), 7.47 (d, J=8.0 Hz, 2H), 7.37 (d, J=8.0 Hz, 2H), 4.82 (m, 1H), 4.12(m, 1H), 3.94 (m, 2H), 3.73 (m, 2H), 3.51 (m, 2H), 3.22 (dd, J=13.2 Hz,J=6.0 Hz, 1H), 3.15 (dd, J=13.2 Hz, J=9.2 Hz, 1H), 2.87 (m, 1H). LCMS(APCI+) m/z 384, 386 [M+H]⁺; Rt=1.92 minutes.

Example 35

Preparation of(2S)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-5-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1:(2S)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting (D)-Boc-4-chlorophenylalanine with(L)-Boc-4-chlorophenylalanine. ¹H NMR (CDCl₃, 400 MHz) δ 10.03 (s, 1H),7.98 (s, 1H), 7.41 (d, J=8.8 Hz, 1H), 7.28 (d, J=8.4 Hz, 2H), 7.16 (d,J=8.4 Hz, 2H), 7.12 (dd, J=8.8 Hz, J=2.0 Hz, 1H), 7.07 (s, 1H), 5.46 (m,1H), 4.88 (m, 1H), 3.74 (m, 2H), 3.53 (m, 1H), 3.31 (m, 1H), 3.07 (m,1H), 2.99 (d, J=6.8 Hz, 2H), 2.91 (m, 2H), 2.49 (m, 1H), 1.43 (s, 9H).

Step 2:(2S)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-5-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2S)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.24 (s, 1H), 8.03 (s,1H), 7.75 (d, J=9.2 Hz, 1H), 7.65 (d, J=9.2 Hz, 1H), 7.47 (d, J=8.0 Hz,2H), 7.37 (d, J=8.0 Hz, 2H), 4.82 (m, 1H), 4.12 (m, 1H), 3.94 (m, 2H),3.73 (m, 2H), 3.51 (m, 2H), 3.22 (dd, J=13.2 Hz, J=6.0 Hz, 1H), 3.15(dd, J=13.2 Hz, J=9.2 Hz, 1H), 2.87 (m, 1H). LCMS (APCI+) m/z 384, 386[M+H]⁺; Rt=1.92 minutes.

Example 36

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-6-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: 6-piperazin-1-yl-1H-indazole was prepared by the proceduresdescribed in Example 34, Step 1, substituting 5-aminoindazole with6-aminoindazole. LCMS (APCI+) m/z 203 [M+H]⁺; Rt=1.53 minutes.

Step 2:(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-6-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with6-piperazin-1-yl-1H-indazole. ¹H NMR (CDCl₃, 400 MHz) δ 9.91 (s, 1H),7.94 (d, J=2.0 Hz, 1H), 7.61 (dd, J=8.8 Hz, J=2.0 Hz, 1H), 7.28 (d,J=8.4 Hz, 2H), 7.16 (d, J=8.4 Hz, 2H), 6.86 (dd, J=8.8 Hz, J=2.0 Hz,1H), 6.73 (s, 1H), 5.42 (m, 1H), 4.88 (m, 1H), 3.74 (m, 2H), 3.53 (m,1H), 3.29 (m, 1H), 3.19 (m, 1H), 3.00 (m, 4H), 2.59 (m, 1H), 1.43 (s,9H). LCMS (APCI+) m/z 484, 486 [M+H]⁺; Rt=3.40 minutes.

Step 3:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-indazol-6-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-6-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.30 (s, 1H), 7.78 (d,J=9.2 Hz, 1H), 7.40 (d, J=8.4 Hz, 2H), 7.32 (d, J=8.4 Hz, 2H), 7.15 (d,J=9.2 Hz, 1H), 6.97 (s, 1H), 4.76 (m, 1H), 3.80 (m, 2H), 3.62 (m, 1H),3.40 (m, 1H), 3.10-3.30 (m, 5H), 2.72 (m, 1H). LCMS (APCI+) m/z 384, 386[M+H]⁺; Rt=2.02 minutes.

Example 37

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1-methyl-1H-indazol-5-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a solution of 5-piperazin-1-yl-1H-indazole (0.34 g, 1.3 mmol)in 1,4-dioxane (5 mL) was added 3N NaOH (0.42 mL, 1.3 mmol). Aftercooling to 0° C., a solution of tert-butylcarbonate (0.25 g, 1.3 mmol)in 1,4-dioxane (1 mL) was added dropwise. The reaction mixture wasstirred at room temperature overnight and then poured into water andextracted with EtOAc. The combined organic layers were washed withsaturated aqueous NaHCO₃, water, brine, dried and concentrated. Theresidue was purified by column chromatography (EtOAc:hexanes, 1:1) togive 4-(1H-Indazol-5-yl)-piperazine-1-carboxylic acid tert-butyl ester(0.31 g, 82%) as a white solid. ¹H NMR (CDCl₃, 400 MHz) δ 10.01 (s, 1H),7.80 (s, 1H), 7.42 (d, J=9.2 Hz, 1H), 7.21 (dd, J=9.2 Hz, J=2.0 Hz, 1H),7.16 (d, J=2.0 Hz, 1H), 3.62 (m, 4H), 3.09 (m, 4H), 1.50 (s, 9H). LCMS(APCI+) m/z 303 [M+H]⁺; Rt=2.50 minutes.

Step 2: To a stirred suspension of NaH (60%, 4 mg, 0.1 mmol) in DMF (0.5mL) was added dropwise a solution of4-(1H-Indazol-5-yl)-piperazine-1-carboxylic acid tert-butyl ester (0.100g, 0.33 mmol) in DMF (1 mL). After stirring for 30 minutes, methyliodide (0.026 g, 0.18 mmol) was added dropwise. The mixture was stirredat room temperature for 2 hours and then partitioned between EtOAc andwater. The aqueous phase was extracted with EtOAc. The combined organiclayers were washed with brine, dried and concentrated. The residue waspurified by column chromatography (hexanes:EtOAc, 2:1) to give4-(1-Methyl-1H-indazol-5-yl)-piperazine-1-carboxylic acid tert-butylester (0.023 g, 22%) as a white solid. ¹H NMR (CDCl₃, 400 MHz) δ 7.87(s, 1H), 7.32 (d, J=9.2 Hz, 1H), 7.20 (d, J=9.2 Hz, 1H), 7.12 (s, 1H),4.04 (s, 3H), 3.62 (m, 4H), 3.09 (m, 4H), 1.50 (s, 9H). LCMS (APCI+) m/z317 [M+H]⁺; Rt=3.31 minutes.

Step 3: 1-Methyl-5-piperazin-1-yl-1H-indazole dihydrochloride wasprepared by the procedures described in Example 34, Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with4-(1-Methyl-1H-indazol-5-yl)-piperazine-1-carboxylic acid tert-butylester. LCMS (APCI+) m/z 217 [M+H]⁺; Rt=1.15 minutes.

Step 4:(2R)-{1-(4-chlorobenzyl)-2-[4-(1-methyl-1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with1-Methyl-5-piperazin-1-yl-1H-indazole dihydrochloride. ¹H NMR (CDCl₃,400 MHz) δ 7.87 (s, 1H), 7.31 (d, J=9.2 Hz, 1H), 7.28 (d, J=8.0 Hz, 2H),7.17 (d, J=8.0 Hz, 2H), 7.12 (d, J=9.2 Hz, 1H), 7.04 (s, 1H), 5.41 (m,1H), 4.87 (m, 1H), 4.04 (s, 3H), 3.73 (m, 2H), 3.50 (m, 1H), 3.31 (m,1H), 3.07 (m, 1H), 2.99 (d, J=7.2 Hz, 2H), 2.94 (m, 2H), 2.48 (m, 1H),1.43 (s, 9H). LCMS (APCI+) m/z 498, 500 [M+H]⁺; Rt=3.27 minutes.

Step 5:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1-methyl-1H-indazol-5-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(1-methyl-1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.13 (s, 1H), 7.96 (s,1H), 7.76 (d, J=9.2 Hz, 1H), 7.62 (dd, J=9.2 Hz, J=2.0 Hz, 1H), 7.47 (d,J=8.4 Hz, 2H), 7.37 (d, J=8.4 Hz, 2H), 4.81 (m, 1H), 4.11 (m, 4H), 3.98(m, 1H), 3.90 (m, 1H), 3.69 (m, 2H), 3.49 (m, 2H), 3.16 (m, 2H), 2.84(m, 1H). LCMS (APCI+) m/z 398, 400 [M+H]⁺; Rt=2.05 minutes.

Example 38

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-isoquinolin-6-yl-piperazin-1-yl)-propan-1-onedihydrochloride

Step 1: A round bottom flask charged with 6-bromo isoquinoline (preparedfrom 4-bromobenzaldehyde according to the literature: Neiko Nerenz, etal. (1998) J. Chem. Soc. Perkin Trans. 2, 437-447, 0.200 g, 0.961 mmol),1-Boc piperazine (0.215 g, 1.15 mmol), K₃PO₄ (0.286 g, 1.35 mmol),(2′-dicyclohexylphosphanyl-biphenyl-2-yl)-dimethylamine (0.028 g, 0.072mmol) and Pd₂ dba₃ (0.022 g, 0.024 mmol) in dry DME (2 mL) was purgedunder N₂ and heated at reflux for 5 hours. After cooling, the mixturewas partitioned between EtOAc and H₂O. The organic layer was washed withbrine, dried and concentrated. The residue was purified by columnchromatography (1:1 hexanes/EtOAc,) to give4-Isoquinolin-6-yl-piperazine-1-carboxylic acid tert-butyl ester (0.210g, 70%) as a white solid. ¹H NMR (CDCl₃, 400 MHz) δ 9.04 (s, 1H), 8.39(dd, J=6.8 Hz, J=2.8 Hz, 1H), 7.83 (dd, J=9.2 Hz, J=2.8 Hz, 1H), 7.45(d, J=6.8 Hz, 1H), 7.32 (dd, J=9.2 Hz, J=2.4 Hz, 1H), 6.98 (s, 1H), 3.64(m, 4H), 3.35 (m, 4H), 1.50 (s, 9H). LCMS (APCI+) m/z 314 [M+H]⁺;Rt=2.14 minutes.

Step 2: 6-piperazin-1-yl-isoquinoline dihydrochloride was prepared bythe procedures described in Example 34, Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with 4-Isoquinolin-6-yl-piperazine-1-carboxylicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 9.28 (s, 1H), 8.30 (d,J=9.2 Hz, 1H), 8.24 (d, J=7.2 Hz, 1H), 8.04 (d, J=6.8 Hz, 1H), 7.83 (dd,J=9.2 Hz, J=2.4 Hz, 1H), 7.49 (s, 1H), 3.98 (m, 4H), 3.45 (m, 4H). LCMS(APCI+) m/z 214 [M+H]⁺; Rt=1.76 minutes.

Step 3:(2R)-[1-(4-chlorobenzyl)-2-(4-isoquinolin-6-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with6-piperazin-1-yl-isoquinoline dihydrochloride. ¹H NMR (CDCl₃, 400 MHz) δ9.04 (s, 1H), 8.40 (dd, J=5.6 Hz, J=3.2 Hz, 1H), 7.83 (dd, J=9.2 Hz,J=2.4 Hz, 1H), 7.46 (d, J=6.0 Hz, 1H), 7.27 (m, 3H), 7.17 (d, J=8.0 Hz,2H), 6.90 (s, 1H), 5.41 (m, 1H), 4.87 (m, 1H), 3.76 (m, 2H), 3.58 (m,1H), 3.31 (m, 2H), 3.20 (m, 2H), 2.99 (d, J=6.8 Hz, 2H), 2.78 (m, 1H),1.43 (s, 9H). LCMS (APCI+) m/z 495, 497 [M+H]⁺; Rt=2.50 minutes.

Step 4:(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-isoquinolin-6-yl-piperazin-1-yl)-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-[1-(4-chlorobenzyl)-2-(4-isoquinolin-6-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 9.18 (s, 1H), 8.22 (d,J=9.2 Hz, 1H), 8.16 (d, J=6.8 Hz, 1H), 7.95 (d, J=6.8 Hz, 1H), 7.69 (dd,J=9.2 Hz, J=2.4 Hz, 1H), 7.38 (d, J=8.4 Hz, 2H), 7.32 (d, J=8.4 Hz, 2H),7.27 (s, 1H), 4.75 (m, 1H), 3.58-3.84 (m, 6H), 3.10-3.30 (m, 4H). LCMS(APCI+) m/z 395, 397 [M+H]⁺; Rt=2.68 minutes.

Example 39

Preparation of(2R)-2-Amino-3-(1H-indol-3-yl)-1-(4-isoquinolin-6-yl-piperazin-1-yl)-propan-1-onedihydrochloride

Step 1:(2R)-[1-(1H-Indol-3-ylmethyl)-2-(4-isoquinolin-6-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with6-piperazin-1-yl-isoquinoline dihydrochloride and substituting(D)-Boc-4-chlorophenylalanine with (D)-Boc-tryptophan. ¹H NMR (CDCl₃,400 MHz) δ 9.00 (s, 1H), 8.37 (d, J=6.4 Hz, 1H), 8.19 (s, 1H), 7.76 (d,J=9.2 Hz, 1H), 7.72 (d, J=7.6 Hz, 1H), 7.41 (d, J=6.0 Hz, 1H), 7.30 (d,J=7.6 Hz, 1H), 7.18 (m, 2H), 7.12 (d, J=8.8 Hz, 1H), 7.08 (s, 1H), 6.70(s, 1H), 5.53 (d, J=8.4 Hz, 1H), 5.03 (m, 1H), 3.79 (m, 1H), 3.47 (m,1H), 3.05-3.40 (m, 6H), 2.73 (m, 1H), 1.97 (m, 1H), 1.46 (s, 9H). LCMS(APCI+) m/z 500 [M+H]⁺; Rt=2.75 minutes.

Step 2:(2R)-2-Amino-3-(1H-indol-3-yl)-1-(4-isoquinolin-6-yl-piperazin-1-yl)-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-[1-(1H-Indol-3-ylmethyl)-2-(4-isoquinolin-6-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 10.65 (s, 1H), 9.14 (s,1H), 8.15 (m, 2H), 7.89 (d, J=6.8 Hz, 1H), 7.56 (d, J=5.6 Hz, 1H), 7.51(d, J=9.2 Hz, 1H), 7.30 (m, 2H), 7.07 (m, 3H), 4.70 (m, 1H), 3.74 (m,1H), 3.64 (m, 1H), 3.39 (m, 5H), 3.01 (m, 1H), 2.86 (m, 1H), 2.36 (m,1H). LCMS (APCI+) m/z 400 [M+H]⁺; Rt=1.73 minutes.

Example 40

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methylsulfanylpyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: A mixture of 4-chloro-5-iodopyrimidine (3.00 g, 12.5 mmol)(prepared from 4(3H)-pyrimidinone according to the literature: TakaoSakamoto, et al. (1986) Chem. Pharm. Bull., 2719-2724), Et₃N (5.22 mL,37.4 mmol), 1-Boc piperazine (2.79 g, 15.0 mmol) and NMP (30 mL) washeated at 75° C. for 6 hours. After cooling, the reaction mixture waspartitioned between EtOAc and water. The aqueous phase was extractedwith EtOAc. The combined organic layers were washed with brine, driedand concentrated. The residue was purified by column chromatography (2:1hexanes/EtOAc) to give 4-(5-Iodo-pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (4.81 g, 99%) as a white solid. ¹H NMR (CDCl₃, 400MHz) δ 8.70 (s, 1H), 8.62 (s, 1H), 3.57 (s, 8H), 1.49 (s, 9H). LCMS(APCI+) m/z 391 [M+H]⁺; Rt=2.96 minutes.

Step 2: 5-Iodo-4-piperazin-1-yl-pyrimidine dihydrochloride was preparedby the procedures described in Example 34, Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with4-(5-Iodo-pyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butyl ester.¹H NMR (CD₃OD, 400 MHz) δ 8.87 (s, 1H), 8.84 (s, 1H), 4.36 (t, J=4.2 Hz,4H), 3.47 (t, J=4.2 Hz, 4H). LCMS (APCI+) m/z 291 [M+H]⁺; Rt=1.45minutes.

Step 3:(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with5-Iodo-4-piperazin-1-yl-pyrimidine dihydrochloride. ¹H NMR (CDCl₃, 400MHz) δ 8.70 (s, 1H), 8.62 (s, 1H), 7.27 (d, J=8.0 Hz, 2H), 7.15 (d,J=8.0 Hz, 2H), 5.38 (m, 1H), 4.84 (m, 1H), 3.73 (m, 2H), 3.65 (m, 1H),3.53 (m, 2H), 3.45 (m, 2H), 3.24 (m, 1H), 3.12 (m, 1H), 2.98 (m, 2H),1.42 (s, 9H). LCMS (APCI+) m/z 572, 574 [M+H]⁺; Rt=3.25 minutes.

Step 4: A round bottom flask was charged with sodium methanethiolate (17mg, 0.25 mmol),(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (71 mg, 0.12 mmol) and2,2,6,6-tetramethyl-heptane-3,5-dione (6 mg, 0.25 equivalents). Aftervacuum purging and refilling of N₂, NMP (3 mL) and CuCl (6 mg, 0.06mmol) was added to this mixture. The reaction was stirred at 130° C. for1 hour. After cooling, the reaction mixture was diluted with EtOAc andfiltered through Celite. The filtrate was washed with brine, dried overNa₂SO₄, filtered and concentrated in vacuo. The residue was purified byflash column chromatography (1:1 hexanes/EtOAc) to give(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methylsulfanylpyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (24 mg, 39%) as a colorless gum. ¹H NMR (CDCl₃,400 MHz) δ 8.56 (s, 1H), 8.29 (s, 1H), 7.26 (d, J=8.4 Hz, 2H), 7.14 (d,J=8.4 Hz, 2H), 5.37 (m, 1H), 4.84 (m, 1H), 3.70 (m, 1H), 3.61 (m, 2H),3.51 (m, 4H), 3.20 (m, 2H), 2.97 (m, 2H), 2.42 (s, 3H), 1.42 (s, 9H).LCMS (APCI+) m/z 492, 494 [M+H]⁺; Rt=3.57 minutes.

Step 5:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methylsulfanylpyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methylsulfanylpyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.64 (s, 1H), 8.30 (s,1H), 7.39 (d, J=8.0 Hz, 2H), 7.32 (d, J=8.0 Hz, 2H), 4.75 (m, 1H), 4.09(m, 2H), 4.00 (m, 1H), 3.83 (m, 1H), 3.68 (m, 3H), 3.17 (m, 3H), 2.55(s, 3H). LCMS (APCI+) m/z 392, 394 [M+H]⁺; Rt=1.85 minutes.

Example 41

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-phenylpyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1:(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (0.150 g, 0.262 mmol), phenylboronic acid (0.042g, 0.341 mmol) and 2N sodium carbonate solution (0.34 mL, 0.68 mmol)were stirred in DME (3 mL) and the mixture was degassed with N₂ for 15minutes. Tetrakis(triphenylphosphine) palladium (0) (0.015 g, 0.013mmol) was added and the mixture heated at 80° C. for 24 hours. Themixture was cooled to room temperature and partitioned between DCM andwater. The aqueous layer was extracted with DCM. The combined organiclayers were washed with saturated aqueous NaHCO₃ and brine, dried andconcentrated. The residue was purified by flash chromatography on silicagel, eluting with hexanes:EtOAc (3:1 to 1:1) to give(2R)-{1-(4-chlorobenzyl)-2-oxo-2-[4-(5-phenylpyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester (0.074 g, 54%) as a colorless oil. ¹H NMR (CDCl₃,400 MHz) δ 8.65 (s, 1H), 8.21 (s, 1H), 7.46 (m, 2H), 7.38 (m, 2H), 7.25(m, 3H), 7.09 (d, J=8.4 Hz, 2H), 5.30 (m, 1H), 4.73 (m, 1H), 3.50 (m,1H), 3.43 (m, 1H), 3.30 (m, 2H), 3.16 (m, 2H), 2.97 (m, 1H), 2.91 (d,J=7.6 Hz, 2H), 2.85 (m, 1H), 1.39 (s, 9H). LCMS (APCI+) m/z 522, 524[M+H]⁺; Rt=2.72 minutes.

Step 2:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-phenylpyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-oxo-2-[4-(5-phenylpyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.75 (s, 1H), 8.20 (s,1H), 7.56 (m, 3H), 7.44 (d, J=6.8 Hz, 2H), 7.37 (d, J=8.0 Hz, 2H), 7.25(d, J=8.0 Hz, 2H), 4.62 (m, 1H), 3.47-3.54 (m, 6H), 3.38-3.44 (m, 4H).LCMS (APCI+) m/z 422, 424 [M+H]⁺; Rt=2.40 minutes.

Example 42

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-thiophen-3-yl-pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1:(2R)-{1-(4-chlorobenzyl)-2-oxo-2-[4-(5-thiophen-3-yl-pyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 41, Step 1, substituting phenylboronic acid with3-thiopheneboronic acid. ¹H NMR (CDCl₃, 400 MHz) δ 8.64 (s, 1H), 8.27(s, 1H), 7.46 (d, J=4.8 Hz, 1H), 7.33 (s, 1H), 7.26 (d, J=8.4 Hz, 2H),7.16 (d, J=4.8 Hz, 1H), 7.11 (d, J=8.4 Hz, 2H), 5.33 (m, 1H), 4.76 (m,1H), 3.55 (m, 1H), 3.48 (m, 1H), 3.27 (m, 2H), 3.16 (m, 2H), 2.98 (m,1H), 2.92 (d, J=6.8 Hz, 2H), 2.82 (m, 1H), 1.40 (s, 9H). LCMS (APCI+)m/z 528, 530 [M+H]⁺; Rt=2.75 minutes.

Step 2:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-thiophen-3-yl-pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-oxo-2-[4-(5-thiophen-3-yl-pyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.74 (s, 1H), 8.24 (s,1H), 7.70 (s, 1H), 7.65 (s, 1H), 7.39 (d, J=8.0 Hz, 2H), 7.27 (d, J=8.0Hz, 2H), 7.22 (d, J=4.8 Hz, 1H), 4.62 (m, 1H), 3.48-3.55 (m, 6H),3.38-3.44 (m, 4H). LCMS (APCI+) m/z 428, 430 [M+H]⁺; Rt=2.76 minutes.

Example 43

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-phenoxypyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

A round bottom flask was charged with phenol (33 mg, 0.35 mmol),(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (100 mg, 0.175 mmol), Cs₂CO₃ (114 mg, 0.350 mmol)and 2,2,6,6-tetramethyl-heptane-3,5-dione (8 mg, 0.25 equivalents).After vacuum purging and refilling of N₂, NMP (3 mL) and CuCl (9 mg,0.09 mmol) was added to this mixture. The reaction was stirred at 130°C. for 2 hours. After cooling, the reaction was diluted with EtOAc andfiltered. The filtrate was washed with water, brine and dried.Evaporation of the solvent followed by flash column chromatography (10:1DCM/MeOH) gave the title compound as a free base, which was converted todihydrochloride salt (36 mg, 38%) by treatment with 4 N HCl in dioxane.¹H NMR (CD₃OD, 400 MHz) δ 8.64 (s, 1H), 7.96 (s, 1H), 7.70 (s, 1H), 7.65(s, 1H), 7.49 (m, 2H), 7.37 (m, 2H), 7.29 (m, 3H), 7.14 (d, J=8.0 Hz,2H), 4.70 (m, 1H), 4.11 (m, 2H), 4.00 (m, 1H), 3.76 (m, 2H), 3.57 (m,2H), 3.10 (m, 3H). LCMS (APCI+) m/z 438, 440 [M+H]⁺; Rt=2.13 minutes.

Example 44

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

The title compound was prepared by the procedures described in Example34, Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.77 (s, 1H), 8.73 (s,1H), 7.41 (d, J=8.4 Hz, 2H), 7.33 (d, J=8.4 Hz, 2H), 4.74 (m, 1H), 4.05(m, 2H), 3.95 (m, 1H), 3.84 (m, 1H), 3.69 (m, 3H), 3.22 (m, 1H), 3.13(m, 2H). LCMS (APCI+) m/z 472, 474 [M+H]⁺; Rt=2.06 minutes.

Example 45

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-chloropyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1:(2R)-{1-(4-chlorobenzyl)-2-[4-(5-chloropyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 40, Step 1, substituting 4-chloro-5-iodopyrimidine with4,5-dichloropyrimidine (prepared from 5-chloropyrimidin-4-ol accordingto the literature: Chestfield J. et al. (1955) J. Chem. Soc. Abstracts,3478-3481), and substituting 1-Boc piperazine with[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acidtert-butyl ester. ¹H NMR (CDCl₃, 400 MHz) δ 8.57 (s, 1H), 8.32 (s, 1H),7.27 (d, J=8.0 Hz, 2H), 7.15 (d, J=8.0 Hz, 2H), 5.36 (m, 1H), 4.85 (m,1H), 3.50-3.80 (m, 6H), 3.24 (m, 2H), 2.98 (m, 2H), 1.43 (s, 9H). LCMS(APCI+) m/z 480, 482, 484 [M+H]⁺; Rt=3.24 minutes.

Step 2:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-chloropyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(5-chloropyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.74 (s, 1H), 8.57 (s,1H), 7.40 (d, J=7.6 Hz, 2H), 7.33 (d, J=7.6 Hz, 2H), 4.74 (m, 1H), 4.13(m, 2H), 4.03 (m, 1H), 3.85 (m, 1H), 3.72 (m, 3H), 3.22 (m, 1H), 3.16(m, 2H). LCMS (APCI+) m/z 380, 382, 384 [M+H]⁺; Rt=2.01 minutes.

Example 46

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-fluoropyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1:(2R)-{1-(4-chlorobenzyl)-2-[4-(5-fluoropyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 40, Step 1, substituting 4-chloro-5-iodopyrimidine with4-chloro-5-fluoropyrimidine (prepared from 5-fluoropyrimidin-4-olaccording to the literature (Kheifets, G. M. et al., 2000, Russian. Org.Chem., 1373-1387), and substituting 1-Boc piperazine with[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acidtert-butyl ester. ¹H NMR (CDCl₃, 400 MHz) δ 8.41 (d, J=2.8 Hz, 1H), 8.12(d, J=6.4 Hz, 1H), 7.27 (d, J=8.0 Hz, 2H), 7.15 (d, J=8.0 Hz, 2H), 5.35(m, 1H), 4.83 (m, 1H), 3.45-3.75 (m, 6H), 3.35 (m, 1H), 3.18 (m, 1H),2.98 (d, J=7.2 Hz, 2H), 1.43 (s, 9H). LCMS (APCI+) m/z 464, 466 [M+H]⁺;Rt=2.93 minutes.

Step 2:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-fluoropyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(5-fluoropyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.66 (s, 1H), 8.50 (s,1H), 7.39 (d, J=8.0 Hz, 2H), 7.32 (d, J=8.0 Hz, 2H), 4.75 (m, 1H), 4.05(m, 2H), 3.97 (m, 1H), 3.88 (m, 1H), 3.70 (m, 3H), 3.23 (m, 1H), 3.15(m, 2H). LCMS (APCI+) m/z 364, 366 [M+H]⁺; Rt=1.88 minutes.

Example 47

Preparation of(2R)-2-Amino-1-[4-(5-benzyloxypyrimidin-4-yl)-piperazin-1-yl]-3-(4-chlorophenyl)-propan-1-onedihydrochloride

Step 1: A sealed tube charged with4-(5-Iodo-pyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butyl ester(780 mg, 2.00 mmol), CuI (38 mg, 0.20 mmol), 1,10-phenathroline (72 mg,0.4 mmol), Cs₂CO₃ (912 mg, 2.8 mmol), benzyl alcohol (0.62 mL, 6.0 mmol)and toluene (2 mL) was heated at 110° C. for 40 hours. The resultingsuspension was cooled to room temperature and filtered through a silicagel pad, eluting with EtOAc. Evaporation of the solvent followed byflash chromatography on silica gel (10:1 hexanes/EtOAc) provided4-(5-benzyloxypyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butylester (0.640 g, 86%) as a white solid. ¹H NMR (CDCl₃, 400 MHz) δ 8.36(s, 1H), 7.99 (s, 1H), 7.39 (m, 5H), 5.08 (s, 2H), 3.73 (m, 4H), 3.48(m, 4H), 1.47 (s, 9H). LCMS (APCI+) m/z 371 [M+H]⁺; Rt=2.52 minutes.

Step 2: 5-Benzyloxy-4-piperazin-1-yl-pyrimidine dihydrochloride wasprepared by the procedures described in Example 34, Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with4-(5-Benzyloxypyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butylester. LCMS (APCI+) m/z 271 [M+H]⁺; Rt=1.64 minutes.

Step 3:(2R)-[2-[4-(5-Benzyloxypyrimidin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with5-Benzyloxy-4-piperazin-1-yl-pyrimidine dihydrochloride. ¹H NMR (CDCl₃,400 MHz) δ 8.34 (s, 1H), 8.00 (s, 1H), 7.38 (m, 5H), 7.24 (d, J=8.0 Hz,2H), 7.12 (d, J=8.0 Hz, 2H), 5.37 (d, J=8.8 Hz, 1H), 5.06 (s, 2H), 4.81(m, 1H), 3.64 (m, 5H), 3.44 (m, 1H), 3.34 (m, 1H), 3.13 (m, 1H), 2.96(m, 2H), 1.41 (s, 9H). LCMS (APCI+) m/z 552, 554 [M+H]⁺; Rt=2.79minutes.

Step 4:(2R)-2-Amino-1-[4-(5-benzyloxypyrimidin-4-yl)-piperazin-1-yl]-3-(4-chlorophenyl)-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-[2-[4-(5-benzyloxypyrimidin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.49 (s, 1H), 8.11 (s,1H), 7.48 (m, 5H), 7.36 (d, J=8.4 Hz, 2H), 7.28 (d, J=8.4 Hz, 2H), 5.20(s, 2H), 4.68 (m, 1H), 4.07 (m, 2H), 3.97 (m, 1H), 3.69 (m, 2H), 3.59(m, 2H), 3.12 (m, 3H). LCMS (APCI+) m/z 452, 454 [M+H]⁺; Rt=2.02minutes.

Example 48

Preparation of(2R)-2-Amino-1-[4-(5-aminopyrimidin-4-yl)-piperazin-1-yl]-3-(4-chlorophenyl)-propan-1-onedihydrochloride

Step 1: 4-piperazin-1-yl-pyrimidin-5-ylamine dihydrochloride wasprepared by the procedures described in Example 34, Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with4-(5-aminopyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butyl ester(prepared from 4,6-dichloro-nitropyrimidine according to the proceduresdescribed in U.S. Pat. No. 5,563,142). LCMS (APCI+) m/z 180 [M+H]⁺;Rt=1.12 minutes.

Step 2:(2R)-[2-[4-(5-aminopyrimidin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with4-piperazin-1-yl-pyrimidin-5-ylamine dihydrochloride. ¹H NMR (CDCl₃, 400MHz) δ 8.38 (s, 1H), 7.97 (s, 1H), 7.27 (d, J=8.0 Hz, 2H), 7.14 (d,J=8.0 Hz, 2H), 5.38 (d, J=8.4 Hz, 1H), 4.85 (m, 1H), 3.66 (m, 2H), 3.52(m, 2H), 3.44 (s, 2H), 3.23 (m, 4H), 2.93 (m, 2H), 1.42 (s, 9H). LCMS(APCI+) m/z 461, 463 [M+H]⁺; Rt=2.38 minutes.

Step 3:(2R)-2-Amino-1-[4-(5-aminopyrimidin-4-yl)-piperazin-1-yl]-3-(4-chlorophenyl)-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-[2-[4-(5-aminopyrimidin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.41 (s, 1H), 7.80 (s,1H), 7.40 (d, J=8.4 Hz, 2H), 7.32 (d, J=8.4 Hz, 2H), 4.77 (m, 1H), 3.84(m, 4H), 3.63 (m, 2H), 3.50 (m, 1H), 3.18 (m, 3H). LCMS (APCI+) m/z 361,363 [M+H]⁺; Rt=1.68 minutes.

Example 49

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methoxypyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a stirred solution of4-(5-benzyloxypyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butylester (0.540 g, 1.46 mmol) in MeOH (20 mL) under N₂ was cautiously added10% Pd on carbon (40 mg). The reaction vessel was evacuated under vacuumand then put under an atmosphere of hydrogen using a balloon. Themixture was stirred for 2 hours at room temperature. At this time thehydrogen gas was evacuated and the catalyst was removed by filtration.The filtrate was concentrated. The residue was purified by flashchromatography (1:1 hexanes/EtOAc) to give4-(5-Hydroxypyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butylester (0.390 g, 95%) as a white solid. ¹H NMR (CDCl₃, 400 MHz) δ 8.18(s, 1H), 7.75 (s, 1H), 3.93 (m, 4H), 3.54 (m, 4H), 1.41 (s, 9H). LCMS(APCI+) m/z 281 [M+H]⁺; Rt=2.01 minutes.

Step 2: 4-piperazin-1-yl-pyrimidin-5-ol dihydrochloride was prepared bythe procedures described in Example 34, Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with4-(5-Hydroxypyrimidin-4-yl)-piperazine-1-carboxylic acid tert-butylester. LCMS (APCI+) m/z 181 [M+H]⁺; Rt=1.15 minutes.

Step 3:(2R)-{1-(4-chlorobenzyl)-2-[4-(5-hydroxypyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with4-piperazin-1-yl-pyrimidin-5-ol dihydrochloride. ¹H NMR (CDCl₃, 400 MHz)δ 8.18 (s, 1H), 7.76 (s, 1H), 7.25 (d, J=8.0 Hz, 2H), 7.14 (d, J=8.0 Hz,2H), 5.39 (d, J=8.4 Hz, 1H), 4.84 (m, 1H), 3.55-3.95 (m, 5H), 3.51 (m,2H), 3.21 (m, 1H), 2.96 (m, 2H), 1.42 (s, 9H). LCMS (APCI+) m/z 462, 464[M+H]⁺; Rt=2.41 minutes.

A mixture of methyl iodide (18 mg, 0.13 mmol), K₂CO₃ (18 mg, 0.13 mmol)and(2R)-{1-(4-chlorobenzyl)-2-[4-(5-hydroxypyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (50 mg, 0.11 mmol) in DMF (2 mL) was stirred atroom temperature for 1 hour. The reaction mixture was diluted with waterand extracted with EtOAc. The combined organic layers were washed withbrine, dried and concentrated. The residue was purified by flashchromatography (hexanes:EtOAc, 1:1) to give(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methoxypyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (20 mg, 39%) as a colorless oil. ¹H NMR (CDCl₃,400 MHz) δ 8.34 (s, 1H), 7.94 (s, 1H), 7.26 (d, J=8.4 Hz, 2H), 7.14 (d,J=8.4 Hz, 2H), 5.37 (d, J=8.4 Hz, 1H), 4.84 (m, 1H), 3.87 (s, 3H), 3.65(m, 5H), 3.48 (m, 1H), 3.33 (m, 1H), 3.20 (m, 1H), 2.96 (m, 2H), 1.42(s, 9H). LCMS (APCI+) m/z 476, 478 [M+H]⁺; Rt=2.44 minutes.

(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methoxypyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methoxypyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester ¹H NMR (CD₃OD, 400 MHz) δ 8.50 (s, 1H), 8.01 (s,1H), 7.38 (d, J=8.0 Hz, 2H), 7.32 (d, J=8.0 Hz, 2H), 4.75 (m, 1H), 4.12(m, 2H), 4.04 (m, 1H), 3.97 (s, 3H), 3.81 (m, 2H), 3.67 (m, 2H), 3.16(m, 3H). LCMS (APCI+) m/z 376, 378 [M+H]⁺; Rt=1.77 minutes.

Example 50

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-isopropoxypyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1:(2R)-{1-(4-chlorobenzyl)-2-[4-(5-isopropoxypyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 49, Step 2, substituting methyl iodide with isopropyl bromide.¹H NMR (CDCl₃, 400 MHz) δ 8.33 (s, 1H), 7.94 (s, 1H), 7.26 (d, J=8.0 Hz,2H), 7.14 (d, J=8.0 Hz, 2H), 5.39 (d, J=8.8 Hz, 1H), 4.84 (m, 1H), 4.48(m, 1H), 3.63 (m, 5H), 3.48 (m, 1H), 3.33 (m, 1H), 3.14 (m, 1H), 2.97(m, 2H), 1.42 (s, 9H), 1.34 (d, J=6.4 Hz, 6H). LCMS (APCI+) m/z 504, 506[M+H]⁺; Rt=2.61 minutes.

Step 2:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-isopropoxypyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(5-isopropoxypyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.47 (s, 1H), 8.03 (s,1H), 7.39 (d, J=8.0 Hz, 2H), 7.33 (d, J=8.0 Hz, 2H), 4.73 (m, 2H), 4.13(m, 2H), 4.02 (m, 1H), 3.81 (m, 2H), 3.66 (m, 2H), 3.17 (m, 3H), 1.42(d, J=6.4 Hz, 6H). LCMS (APCI+) m/z 404, 406 [M+H]⁺; Rt=1.86 minutes.

Example 51

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methylpyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: A mixture of(2R)-{1-(4-chlorobenzyl)-2-[4-(5-iodopyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (0.070 g, 0.12 mmol), methylboronic acid (0.022 g,0.37 mmol), K₂CO₃ (0.085 g, 0.61 mmol) and PdCl₂(PPh₃)₂ (0.0086 g, 0.012mmol) in DMF (2 mL) was heated at 100° C. for 16 hours under nitrogen.The mixture was cooled to room temperature and partitioned between EtOAcand water. The organic layer was washed with saturated aqueous NaHCO₃and brine, dried and concentrated. The residue was purified by flashchromatography on silica gel, eluting with DCM/MeOH (70:1) to give(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methylpyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (0.022 g, 39%) as a colorless oil. ¹H NMR (CDCl₃,400 MHz) δ 8.62 (s, 1H), 8.20 (s, 1H), 7.26 (d, J=8.0 Hz, 2H), 7.14 (d,J=8.0 Hz, 2H), 5.37 (d, J=8.4 Hz, 1H), 4.84 (m, 1H), 3.70 (m, 1H), 3.60(m, 1H), 3.42 (m, 1H), 3.32 (m, 4H), 3.02 (m, 3H), 2.20 (s, 3H), 1.42(s, 9H). LCMS (APCI+) m/z 460, 462 [M+H]⁺; Rt=2.38 minutes.

Step 2:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methylpyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(5-methylpyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.65 (s, 1H), 8.13 (s,1H), 7.39 (d, J=8.4 Hz, 2H), 7.32 (d, J=8.4 Hz, 2H), 4.73 (m, 1H), 3.40(m, 2H), 3.96 (m, 1H), 3.80 (m, 1H), 3.67 (m, 3H), 3.21 (m, 1H), 3.14(m, 2H). LCMS (APCI+) m/z 360, 362 [M+H]⁺; Rt=1.70 minutes.

Example 52

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-cinnolin-4-yl-piperazin-1-yl)-propan-1-onedihydrochloride

Step 1: To a suspension of NaH (60% in mineral oil, 0.099 g, 2.46 mmol)in DMF (5 mL) was added cinnolin-4-ol (prepared from 2-aminoacetophenoneaccording to the procedures described in U.S. Pat. No. 4,620,000), 0.300g, 2.05 mmol) in DMF (2 mL) dropwise. The reaction mixture was warmed at40° C. and stirred for 30 minutes. After cooling,N-phenyltrifluoromethanesulfonimide (0.880 g, 2.46 mmol) in DMF (2 mL)was added, and the reaction mixture was stirred at room temperature for1 hour. 1-Boc piperazine (0.765 g, 4.11 mmol) was added to the mixture.The reaction was stirred at 80° C. for 4 hours. After cooling, themixture was partitioned between EtOAc and water. The aqueous phase wasextracted with EtOAc. The combined organic layers were washed withwater, brine, dried and concentrated. The residue was purified by columnchromatography (1:1 to 1:3 hexanes/EtOAc) to give4-Cinnolin-4-yl-piperazine-1-carboxylic acid tert-butyl ester (0.246 g,38%) as a yellow oil. ¹H NMR (CDCl₃, 400 MHz) δ 8.91 (s, 1H), 8.47 (d,J=8.4 Hz, 1H), 7.97 (d, J=8.4 Hz, 1H), 7.80 (t, J=7.2 Hz, 1H), 7.69 (t,J=7.2 Hz, 1H), 3.74 (m, 4H), 3.34 (m, 4H), 1.51 (s, 9H). LCMS (APCI+)m/z 315 [M+H]⁺; Rt=2.14 minutes.

Step 2: 4-piperazin-1-yl-cinnoline dihydrochloride was prepared by theprocedures described in Example 34, Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with 4-Cinnolin-4-yl-piperazine-1-carboxylic acidtert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.96 (s, 1H), 8.33 (d, J=8.8Hz, 1H), 8.15 (t, J=7.2 Hz, 1H), 8.07 (d, J=8.4 Hz, 1H), 7.88 (t, J=7.2Hz, 1H), 4.42 (m, 4H), 3.64 (m, 4H). LCMS (APCI+) m/z 215 [M+H]⁺;Rt=1.46 minutes.

Step 3:(2R)-[1-(4-chlorobenzyl)-2-(4-cinnolin-4-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with4-piperazin-1-yl-cinnoline dihydrochloride. ¹H NMR (CDCl₃, 400 MHz) δ8.82 (s, 1H), 8.48 (d, J=8.4 Hz, 1H), 7.89 (d, J=8.4 Hz, 1H), 7.80 (t,J=7.2 Hz, 1H), 7.70 (t, J=7.2 Hz, 1H), 7.31 (d, J=8.4 Hz, 2H), 7.18 (d,J=8.4 Hz, 2H), 5.36 (m, 1H), 4.88 (m, 1H), 3.85 (m, 2H), 3.69 (m, 1H),3.40 (m, 1H), 3.28 (m, 1H), 3.17 (m, 2H), 3.01 (d, J=7.2 Hz, 2H), 2.78(m, 1H), 1.43 (s, 9H). LCMS (APCI+) m/z 496, 498 [M+H]⁺; Rt=2.44minutes.

Step 4:(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-cinnolin-4-yl-piperazin-1-yl)-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-[1-(4-chlorobenzyl)-2-(4-cinnolin-4-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.75 (s, 1H), 8.27 (d,J=8.4 Hz, 1H), 8.10 (t, J=7.2 Hz, 1H), 7.99 (d, J=8.4 Hz, 1H), 7.82 (t,J=7.2 Hz, 1H), 7.42 (d, J=8.0 Hz, 2H), 7.34 (d, J=8.0 Hz, 2H), 4.74 (m,1H), 4.19 (m, 3H), 3.89 (m, 4H), 3.41 (m, 1H), 3.18 (d, J=7.6 Hz, 2H).LCMS (APCI+) m/z 396, 398 [M+H]⁺; Rt=2.46 minutes.

Example 53

Preparation of(2R)-2-Amino-1-(4-cinnolin-4-yl-piperazin-1-yl)-3-phenylpropan-1-onedihydrochloride

Step 1:(2R)-[1-Benzyl-2-(4-cinnolin-4-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester was prepared by the procedures described inExample 34, Step 2, substituting 5-piperazin-1-yl-1H-indazole with4-piperazin-1-yl-cinnoline dihydrochloride, and substituting(D)-Boc-4-chlorophenylalanine with (D)-Boc-phenylalanine. ¹H NMR (CDCl₃,400 MHz) δ 8.73 (s, 1H), 8.46 (d, J=8.4 Hz, 1H), 7.85 (d, J=8.4 Hz, 1H),7.79 (t, J=8.0 Hz, 1H), 7.68 (t, J=8.0 Hz, 1H), 7.29 (m, 5H), 5.42 (d,J=8.4 Hz, 1H), 4.91 (m, 1H), 3.89 (m, 1H), 3.67 (m, 1H), 3.57 (m, 1H),3.32 (m, 2H), 3.08 (m, 2H), 2.99 (m, 2H), 2.32 (m, 1H), 1.45 (s, 9H).LCMS (APCI+) m/z 462 [M+H]⁺; Rt=2.30 minutes.

Step 2:(2R)-2-Amino-1-(4-cinnolin-4-yl-piperazin-1-yl)-3-phenylpropan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-[1-Benzyl-2-(4-cinnolin-4-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.72 (s, 1H), 8.25 (d,J=8.4 Hz, 1H), 8.09 (t, J=8.0 Hz, 1H), 7.99 (d, J=8.4 Hz, 1H), 7.80 (t,J=8.0 Hz, 1H), 7.37 (m, 2H), 4.75 (m, 1H), 4.24 (m, 1H), 4.12 (m, 2H),3.89 (m, 3H), 3.65 (m, 1H), 3.23 (m, 3H). LCMS (APCI+) m/z 362 [M+H]⁺;Rt=2.38 minutes.

Example 54

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(2-methylquinazolin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1:(2R)-{1-(4-chlorobenzyl)-2-[4-(2-methylquinazolin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 40, Step 1, substituting 4-chloro-5-iodopyrimidine with4-chloro-2-methylquinazoline, and substituting 1-Boc piperazine with[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acidtert-butyl ester. ¹H NMR (CDCl₃, 400 MHz) δ 7.84 (d, J=8.4 Hz, 1H), 7.77(d, J=8.4 Hz, 1H), 7.71 (t, J=8.4 Hz, 1H), 7.41 (t, J=8.4 Hz, 1H), 7.28(d, J=8.4 Hz, 2H), 7.16 (d, J=8.4 Hz, 2H), 5.37 (d, J=8.4 Hz, 1H), 4.87(m, 1H), 3.73 (m, 3H), 3.60 (m, 4H), 3.29 (m, 2H), 2.98 (m, 2H), 2.84(s, 3H), 1.43 (s, 9H). LCMS (APCI+) m/z 510, 512 [M+H]⁺; Rt=2.50minutes.

Step 2:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(2-methylquinazolin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(2-methylquinazolin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.19 (d, J=8.0 Hz, 1H),8.01 (t, J=8.0 Hz, 1H), 7.77 (d, J=8.0 Hz, 1H), 7.72 (t, J=8.0 Hz, 1H),7.40 (d, J=8.4 Hz, 2H), 7.34 (d, J=8.4 Hz, 2H), 4.76 (m, 1H), 4.27 (m,2H), 4.19 (m, 1H), 3.70-3.95 (m, 4H), 3.17 (m, 3H). LCMS (APCI+) m/z410, 412 [M+H]⁺; Rt=1.80 minutes.

Example 55

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(2-chloroquinazolin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1:(2R)-{1-(4-chlorobenzyl)-2-[4-(2-chloroquinazolin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester was prepared by the procedures described inExample 40, Step 1, substituting 4-chloro-5-iodopyrimidine with2,4-dichloroquinazoline, and substituting 1-Boc piperazine with[1-(4-chlorobenzyl)-2-oxo-2-piperazin-1-yl-ethyl]-carbamic acidtert-butyl ester. ¹H NMR (CDCl₃, 400 MHz) δ 7.84 (d, J=8.0 Hz, 1H), 7.77(m, 2H), 7.47 (t, J=8.0 Hz, 1H), 7.29 (d, J=8.4 Hz, 2H), 7.16 (d, J=8.4Hz, 2H), 5.35 (d, J=8.8 Hz, 1H), 4.84 (m, 1H), 3.74 (m, 5H), 3.40 (m,1H), 3.31 (m, 1H), 2.99 (d, J=8.0 Hz, 2H), 1.43 (s, 9H). LCMS (APCI+)m/z 530, 532, 534 [M+H]⁺; Rt=3.74 minutes.

Step 2:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(2-chloroquinazolin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-{1-(4-chlorobenzyl)-2-[4-(2-chloroquinazolin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.16 (d, J=8.4 Hz, 1H),7.97 (t, J=8.4 Hz, 1H), 7.74 (d, J=8.4 Hz, 1H), 7.69 (t, J=8.4 Hz, 1H),7.40 (d, J=8.0 Hz, 2H), 7.33 (d, J=8.0 Hz, 2H), 4.74 (m, 1H), 4.14 (m,2H), 4.06 (m, 1H), 3.91 (m, 1H), 3.78 (m, 3H), 3.33 (m, 1H), 3.17 (m,2H). LCMS (APCI+) m/z 430, 432, 434 [M+H]⁺; Rt=2.24 minutes.

Example 56

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(2-methoxyquinazolin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

To a solution of(2R)-{1-(4-chlorobenzyl)-2-[4-(2-chloroquinazolin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (23 mg, 0.043 mmol) in MeOH (1 mL) was added 4 NHCl in dioxane (1 mL). The mixture was stirred at room temperature for 2days and then evaporated to give the title compound (14 mg, 65%) as awhite solid. ¹H NMR (CD₃OD, 400 MHz) δ 8.17 (d, J=7.2 Hz, 1H), 7.94 (t,J=7.2 Hz, 1H), 7.60 (d, J=8.0 Hz, 2H), 7.39 (m, 4H), 4.75 (m, 1H),4.10-4.35 (m, 6H), 3.86 (m, 4H), 3.17 (m, 3H). LCMS (APCI+) m/z 426, 428[M+H]⁺; Rt=1.86 minutes.

Example 57

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-quinolin-4-yl-piperazin-1-yl)-propan-1-one

Step 1: To a solution of 4-chloroquinoline (2.0 g, 12.2 mmol) in toluene(100 mL) was added piperazine (7.98 g, 92.7 mmol). The reaction mixturewas heated to reflux and stirred for 96 hours, after which it was cooledto room temperature and then further cooled to 0° C. The resultingmixture was filtered to remove the hydrochloride salts that hadprecipitated. After washing the salts with toluene, the combinedfiltrate was washed with 10% aqueous acetic acid (2×25 mL). The combinedaqueous extracts were washed with diethyl ether (25 mL) and thenbasified to pH 8-10 by adding 1M NaOH. The resulting aqueous mixture wasextracted with dichloromethane (3×25 mL). The combined organic extractswere dried (Na₂SO₄), filtered, and concentrated by rotary evaporation.The crude residue, 4-piperazinylquinoline, was obtained as a yellowsolid (2.54 g, 97%) and used without further purification. ¹H NMR wasconsistent with that reported in the literature (Abel, M. D., et al.,Journal of Heterocyclic Chemistry (1996), 33(2), 415-420).

Step 2: To a Jones tube containing PS-CDI (Argonaut, 1.04 mmol/g, 2.2equivalents) suspended in a solution of 4-piperazinylquinoline (1.0equivalent) in CHCl₃ was added the solid Boc-protected amino acid (1.5equivalents.). The reaction mixture was shaken for 15 hours at roomtemperature, after which it was vacuum filtered, the resin rinsed withCHCl₃, and the filtrate concentrated by rotary evaporation. Ifnecessary, the crude coupled product was purified on silica (DCM/EtOAcor DCM/MeOH). The resulting Boc-amino amide was dissolved in minimaldioxane, and 4M HCl/dioxane (10 equivalents) was added. The suspensionwas sonicated 5 minutes and stirred at room temperature for 12 hours,after which it was concentrated by rotary evaporation. The solids weredispersed in ether, isolated by filtration with nitrogen pressure, anddried under reduced pressure to give the corresponding amino amide asthe hydrochloride salt, which was 90% pure by HPLC analysis. R_(t) 2.36.MS (APCI+) [M+H]⁺ 395. R_(t) 2.36. MS (APCI+) [M+H]⁺ 395.

Example 58

Preparation of 1-Quinazolin-4-yl-piperidine-4-carboxylic acid(2-amino-ethyl)-(4-chlorobenzyl)-amide dihydrochloride

Step 1: Triethylamine (12.7 mL, 91.1 mmol) was added to a solution of4-chloro-quinazoline (5.00 g, 30.4 mmol) and ethyl isonipecotate (4.78g, 30.4 mmol) in THF (80 mL) at ambient temperature. After beingrefluxed for 12 hours, the mixture was concentrated in vacuo, and theresulting residue was partitioned between DCM and aqueous 0.1 M NaOH.The separated DCM layer was dried (Na₂SO₄), filtered, and concentratedin vacuo. The resulting solids were suspended in Et₂O and filtered togive 1-quinazolin-4-yl-piperidine-4-carboxylic acid ethyl ester (8.0 g,28.0 mmol). This material was dissolved in EtOH (50 mL) and THF (50 mL)followed by the addition of NaOH (3.73 g, 93.4 mmol) in H₂O (50 mL).After being stirred for 12 hours, the mixture was neutralized with 1.0 NHCl (93.5 mL) and concentrated in vacuo. After the volatile organics areremoved, a white precipitate forms in the resulting aqueous solution.The solid was filtered off and air-dried to give1-quinazolin-4-yl-piperidine-4-carboxylic acid (6.5 g). ¹H NMR (DMSO-d₆,400 MHz) δ 12.28 (bs, 1H), 8.57 (s, 1H), 7.91 (m, 1H), 7.75 (m, 2H),7.49 (m, 1H), 4.13 (m, 2H), 3.22 (m, 2H), 2.57 (m, 1H), 1.93 (m, 2H),1.72 (m, 2H).

Step 2: Thionyl chloride (1.37 mL, 18.8 mmol) was added to a suspensionof 1-quinazolin-4-yl-piperidine-4-carboxylic acid (2.20 g, 8.55 mmol) inDCM (40 mL), which results in a clear solution. After being stirred for2 hours, a precipitate forms and was filtered off to give1-quinazolin-4-yl-piperidine-4-carbonyl chloride hydrochloride as awhite solid (2.0 g).

Step 3: A solution of tert-butyl N-(2-aminoethyl)carbamate (1.00 g, 6.24mmol) and 4-chlorobenzaldehyde (0.90 g, 6.37 mmol) in DCE (10 mL) wasstirred for 30 minutes, followed by the addition of NaBH(OAc)₃ (1.98 g,9.36 mmol) in a single portion. After being stirred for 12 hours, themixture was acidified to pH 2 with 0.2 N HCl, and extracted with DCM (3times, each discarded). The acidic aqueous layer was basified to pH 10with 2.0 M NaOH, and extracted with DCM. The DCM extracts were dried(Na₂SO₄), filtered, and concentrated in vacuo to give[2-(4-chlorobenzylamino)-ethyl]-carbamic acid tert-butyl ester as an oil(1.0 g). LCMS (APCI+) m/z 285, 287 [M+H]⁺.

Step 4: 1-Quinazolin-4-yl-piperidine-4-carbonyl chloride hydrochloride(197 mg, 0.63 mmol) was added to a solution of[2-(4-chlorobenzylamino)-ethyl]-carbamic acid tert-butyl ester (180 mg,0.63 mmol) and DMAP (154 mg, 1.26 mmol) in DCM (6.5 mL) cooled in an icebath. After being stirred for 12 hours, the mixture was partitionedbetween DCM (50 mL) and H₂O (80 mL) containing 1 mL of 1.0 M HCl. TheDCM layer was drained off and the acidic aqueous layer was extracted 3more times with DCM. The combined DCM extracts were dried (Na₂SO₄),filtered, and concentrated in vacuo. The crude material waschromatographed (SiO₂) using EtOAc as eluent to give{2-[(4-chlorobenzyl)-(1-quinazolin-4-yl-piperidine-4-carbonyl)-amino]-ethyl}-carbamicacid tert-butyl ester (190 mg).

Step 5:1{2-[(4-Chlorobenzyl)-(1-quinazolin-4-yl-piperidine-4-carbonyl)-amino]-ethyl}-carbamicacid tert-butyl ester (190 mg, 0.36 mmol) was dissolved in DCM (5 mL)followed by the addition of 2.0 M HCl in Et₂O (2 mL). After beingstirred for 12 hours, the mixture was diluted with DCE and concentratedin vacuo. The resulting white solid was then suspended in MeCN andconcentrated in vacuo (repeated twice) to give1-quinazolin-4-yl-piperidine-4-carboxylic acid(2-amino-ethyl)-(4-chlorobenzyl)-amide dihydrochloride as a white powder(120 mg). ¹H NMR (DMSO-d₆, 400 MHz) δ 8.83 (m, 1H), 8.51 (bs, 2H), 8.20(m, 2H), 8.01 (m, 2H), 7.71 (m, 1H), 7.48 (d, J=8.3 Hz, 1H), 7.39 (d,J=8.3 Hz, 1H), 7.31 (d, J=8.3 Hz, 1H), 7.25 (d, J=8.3 Hz, 1H), 4.79 (m,2H), 4.55 (s, 1H), 3.59 (m, 7H), 3.03 (m, 1H), 2.90 (m, 1H), 1.92 (m,4H). LCMS (APCI+) m/z 424, 426 [M+H].

Example 59

Preparation of2-(R)-Amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: A mixture of 4-chloro-1H-pyrrolo[2,3-b]pyridine (2.50 g, 16.4mmol) and N-benzyl piperazine (3.18 g, 18.0 mmol) were melted at 175° C.for 3 hours in a sealed tube, resulting in the formation of acrystalline solid mass. A solution of 0.1 M aqueous NaOH (10 mL) wasadded and the solid was broken up to give a suspension. Filtration gave4-(4-benzyl-piperazin-1-yl)-1H-pyrrolo[2,3-b]pyridine as a white solid(3.90 g). LCMS (APCI+) m/z 293 [M+H]⁺.

Step 2: A solution of4-(4-benzyl-piperazin-1-yl)-1H-pyrrolo[2,3-b]pyridine (3.90 g, 13.3mmol) and Pd(OH)₂/C (937 mg, 1.33 mmol) in MeOH (60 mL) was stirredunder 1 atmosphere of H₂ for 2 d. The mixture was diluted with MeOH,filtered through diatomaceous earth, and the filtrate was concentratedin vacuo to give 4-piperazin-1-yl-1H-pyrrolo[2,3-b]pyridine as a solid(100 mg kept as free base). The remaining material was suspended in MeOHand treated with 2.0 M HCl in Et₂O. This mixture was concentrated invacuo to give 4-piperazin-1-yl-1H-pyrrolo[2,3-b]pyridine dihydrochloride(2.30 g). ¹H NMR (DMSO-d₆, 400 MHz) δ 11.5 (bs, 1H), 9.43 (bs, 2H), 7.96(d, J=5.4 Hz, 1H), 7.25 (d, J=3.3 Hz, 1H), 6.49 (d, J=3.4 Hz, 1H), 6.45(d, J=5.4 Hz, 1H), 3.56 (bs, 4H), 3.21 (bs, 4H).

Step 3: PyBrop (407 mg, 0.87 mmol) was added in a single portion to asolution of (R)—N-Boc-4-chlorophenylalanine (458 mg, 1.53 mmol) and4-piperazin-1-yl-1H-pyrrolo[2,3-b]pyridine (200 mg, 0.73 mmol) in DCM (5mL) cooled in an ice bath. DIEA (0.66 mL, 3.78 mmol) was then droppedin, the ice bath was removed, and the mixture was stirred for 12 hoursat ambient temperature. The mixture was diluted with DCM and washed with0.1 N HCl. The separated DCM layer was dried (Na₂SO₄), filtered, andconcentrated in vacuo. This material was dissolved in MeOH (4 mL)followed by the addition of LiOH monohydrate (122 mg, 2.91 mmol) in H₂O(2 mL) and stirred for 12 hours. The mixture was concentrated in vacuoand chromatographed (SiO₂) using 2% MeOH/DCM followed by 5% MeOH/DCM aseluent. The resulting material was dissolved in DCM (4 mL) and 2.0 M HClin Et₂O (2 mL), and then stirred for 12 hours. The mixture wasconcentrated in vacuo and chromatographed (SiO₂) using 10% MeOH/DCMfollowed by 10% (7 N NH₃ in MeOH)/DCM as eluent. The purified materialwas dissolved in MeOH followed by the addition of 2.0 N HCl in Et₂O, andthen concentrated in vacuo. The resulting glass was suspended in Et₂OOand concentrated in vacuo (repeat twice) to give2-(R)-amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride as an off-white powder (50 mg). ¹H NMR (DMSO-d₆, 400MHz) δ 12.6 (s, 1H), 8.56 (bs, 3H), 8.03 (d, J=7.1 Hz, 1H), 7.38 (m,3H), 7.32 (m, 2H), 6.84 (s, 1H), 6.69 (d, J=7.1 Hz, 1H), 4.63 (bs, 1H),3.88-3.58 (m, 7H), 3.33 (m, 1H), 3.14 (m, 1H), 3.05 (m, 1H). LCMS(APCI+) m/z 384, 386 [M+H]⁺.

Example 60

Preparation of3-Amino-1-(4-quinazolin-4-yl-piperazin-1-yl)-2-p-tolyl-propan-1-onedihydrochloride

Step 1: n-BuLi (1.60M in hexanes, 40.7 mL, 65.1 mmol) was added to a 0°C. solution of diisopropylamine (9.4 mL, 67.0) in 280 mL THF. Themixture was allowed to stir at 0° C. for 30 minutes, then cooled to −78°C. A solution of p-tolyl-acetic acid methyl ester (10.48 g, 63.8 mmol;prepared from p-tolyl-acetic acid) in 10 mL of THF was added to the −78°C. LDA solution by syringe, which was then stirred for 45 minutes. Neattert-butyl bromoacetate (28 mL) was added by syringe, and the reactionwas stirred 15 minutes at −78° C. The bath was removed, and the reactionwas allowed to warm to room temperature. After stirring an additional 5hours, the reaction mixture was quenched with saturated NH₄Cl solution,and the organics were removed in vacuo. The oily mixture was extractedwith ethyl acetate, and the organics were combined. The organic wasdried over MgSO₄, filtered, and concentrated in vacuo. The crude oil waspurified by on silica gel (25:1 hexanes:EtOAc) to afford the2-p-tolyl-succinic acid 4-tert-butyl ester 1-methyl ester as a paleyellow oil (15.3 g, 86%). ¹H NMR (CDCl₃, 400 MHz) δ 7.16 (d, J=8.0 Hz,2H), 7.12 (d, J=8.0 Hz, 2H), 3.99 (dd, J=10.4, 5.6 Hz, 1H), 3.66 (s,3H), 3.09 (dd, J=16.8, 10.4 Hz, 1H), 2.57 (dd, J=16.8, 5.6 Hz, 1H), 2.32(s, 3H), 1.41 (m, 1H). HPLC R_(t)=3.71 min.

Step 2: A solution of 2-p-tolyl-succinic acid 4-tert-butyl ester1-methyl ester (15.3 g, 54.8 mmol) in 110 mL of DCM was treated withneat TFA (63 mL) at room temperature. The mixture was stirred for fivehours to completion, after which the reaction mixture was concentratedand dried in vacuo overnight to afford a white solid. The solid wassuspended in 190 mL of toluene, cooled to 0° C., and treatedsuccessively with diphenylphosphoryl azide (13.4 mL, 62.1 mmol) andtriethyl amine (19.7 mL, 141 mmol). The reaction mixture (homogeneous)was allowed to warm to room temperature and stirred for four hours tocompletion. The solution was quenched with 1% citric acid solution andextracted with EtOAc. The combined organic was washed with brine, driedover Na₂SO₄, filtered, and concentrated in vacuo to give a light brownoil. The crude azide was dissolved in 190 mL of tert-butanol, treatedwith neat SnCl₄ (0.25 mL, 2.82 mmol), and carefully heated to 90° C.with evolution of nitrogen. The mixture was stirred at 90° C. for 2.5hours and cooled to room temperature. The solution was quenched withsaturated NaHCO₃ solution and then concentrated. The oily mixture wasextracted with EtOAc, and the combined organic was washed with brine,dried over MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by on silica gel (9:1 hexanes:EtOAc) to afford the3-tert-butoxycarbonylamino-2-p-tolyl-propionic acid methyl ester as apale yellow oil (12.3 g, 74%). ¹H NMR (CDCl₃, 400 MHz) δ 7.14 (s, 4H),4.86 (br s, 1H), 3.85 (m, 1H), 3.68 (s, 3H), 3.58 (m, 1H), 3.49 (m, 1H),2.33 (s, 3H), 1.42 (s, 9H). HPLC R_(t)=3.31 min.

Step 3: The 3-tert-butoxycarbonylamino-2-p-tolyl-propionic acid methylester (12.3 g, 41.9 mmol) was dissolved in 200 mL 1:1 THF:water andtreated with lithium hydroxide monohydrate (2.64 g, 62.9 mmol) at roomtemperature. The reaction was stirred at room temperature overnight tocompletion and concentrated in vacuo. The oily mixture was partitionedwith water and washed with EtOAc (discarded). The aqueous was treatedwith solid KHSO₄ until pH<2, then extracted with EtOAc. The combinedorganic was dried over Na₂SO₄, filtered, and concentrated in vacuo toafford the 3-tert-butoxycarbonylamino-2-p-tolyl-propionic acid as awhite solid (10.95 g, 93%). ¹H NMR (ca. 1:1 mixture of rotamers) (CDCl₃,400 MHz) δ 10.40-8.40 (br s, 1H), 7.16 (d, J=8.4 Hz, 2H), 7.13 (d, J=8.4Hz, 2H), 6.80 or 4.91 (br s, 1H), 3.86 and 3.75 (m, 1H), 3.55 (m, 1H),3.47 (m, 2H), 2.31 (s, 3H), 1.44 and 1.41 (s, 9H). LCMS (APCI—) m/z 557[2M−H]⁻. HPLC R_(t)=2.80 min.

Step 4: The 4-piperazin-1-yl-quinazoline dihydrochloride (50 mg, 0.174mmol, free-based with 2N NaOH and extracted with DCM), HOBt monohydrate(27 mg, 0.174 mmol), and 3-tert-butoxycarbonylamino-2-p-tolyl-propionicacid (58 mg, 0.209 mmol) were dissolved in 1.3 mL of DCM/3-5 drops ofTHF. The reaction mixture was treated with DCC (43 mg, 0.209 mmol) andallowed to stir at room temperature for 2.5 hours to completion. Themixture was diluted with DCM, vacuum filtered through compressed Celite,and rinsed with DCM. The filtrate was stirred with 2N sodium hydroxidesolution for five minutes, transferred to seperatory funnel, andextracted with DCM. The combined organic was dried over Na₂SO₄,filtered, and concentrated. The residue was purified on silica gel (1:19DCM:EtOAc) to afford3-Boc-amino-1-(4-quinazolin-4-yl-piperazin-1-yl)-2-p-tolyl-propan-1-one.The material was dissolved in 1.0 mL of 1,4-dioxane and treated with 1.0mL of 4M HCl in 1,4-dioxane (precipitation). The mixture was stirred atroom temperature overnight to completion, then concentrated to dryness.The solid was dissolved in a minimal amount of MeOH, then trituratedwith diethyl ether. The resulting solid was isolated by filtrationthrough a fritted funnel with nitrogen pressure, rinsed with ether, anddried in vacuo to afford the3-amino-1-(4-quinazolin-4-yl-piperazin-1-yl)-2-p-tolyl-propan-1-onedihydrochloride as a pale yellow powder (60 mg, 77%). ¹H NMR (DMSO-d₆,400 MHz) δ 8.84 (s, 1H), 8.16 (d, J=8.4 Hz, 1H), 8.00 (m, 4H), 7.91 (d,J=8.0 Hz, 1H), 7.69 (t, J=8.0 Hz, 1H), 7.20 (s, 4H), 4.40 (m, 1H), 4.14(m, 3H), 3.81 (m, 4H), 3.45 (m, 2H), 2.94 (m, 1H), 2.28 (s, 3H). LCMS(APCI+) m/z 376 [M+H]⁺. HPLC R_(t)=1.67 min.

Example 61

Preparation of4-Amino-2-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-one

Step 1: The (4-chlorophenyl)-acetic acid (20.0 g, 106 mmol) wasdissolved in 220 mL of ethanol at ambient temperature. A catalyticamount of sulfuric acid (10 drops) was added to afford a light yellowsolution. The reaction was allowed to stir overnight to completion andwas concentrated to 30 mL. The concentrate was partitioned between ethylacetate and half-saturated NaHCO₃ solution. The aqueous was extractedwith ethyl acetate, and the organics were combined. The organic waswashed with water, brine, separated, dried over MgSO₄, filtered, andconcentrated in vacuo to afford the desired pure (4-chlorophenyl)-aceticacid ethyl ester as a pale yellow oil (21.0 g, 99%). ¹H NMR (CDCl₃, 400MHz) δ 7.29 (d, J=8.4 Hz, 2H), 7.22 (d, J=8.4 Hz, 2H), 4.15 (q, J=7.2Hz, 2H), 3.58 (s, 2H), 1.25 (t, J=7.2 Hz, 3H).

Step 2: The (4-chlorophenyl)-acetic acid ethyl ester (9.52 g, 47.9 mmol)was dissolved in 80 mL of THF, cooled to 0° C., and treated withpotassium tert-butoxide (538 mg, 4.79 mmol). The resulting orangesolution was allowed to stir for 15 minutes at 0° C., then cooled to−78° C. The tert-butyl acrylate (7.72 mL, 52.7 mmol) was added in threeequal portions over ten minutes. The solution was allowed to stirovernight warming slowly to room temperature. The reaction solution wasconcentrated in vacuo, and the residue was partitioned between ethylacetate and saturated NH₄Cl solution. The aqueous was extracted withethyl acetate, and the organics were combined. The organic was washedwith brine, separated, dried over MgSO₄, filtered, and concentrated invacuo. The residue was purified by chromatography (silica gel elutedwith 9:1 hexanes:EtOAc) to afford the 2-(4-chlorophenyl)-pentanedioicacid 5-tert-butyl ester 1-ethyl ester in greater than 80% purity (9.00g, 57%). ¹H NMR (CDCl₃, 400 MHz) δ 7.31-7.21 (m, 4H), 4.13 (q, J=7.6 Hz,2H), 3.58 (t, J=8.0 Hz, 1H), 2.27 (m, 1H), 2.16 (t, J=7.2 Hz, 2H), 2.04(m, 1H), 1.43 (s, 9H), 1.21 (t, J=7.6 Hz, 3H).

Step 3: The 2-(4-chlorophenyl)-pentanedioic acid 5-tert-butyl ester1-ethyl ester (9.00 g, 27.5 mmol) was dissolved in 40 mL of DCM at roomtemperature and treated slowly with 40 mL of TFA. The solution wasallowed to stir for three hours to completion, then concentrated invacuo. The residue was stored under vacuum overnight then dissolved in80 mL of toluene. The solution was degassed under nitrogen, cooled to 0°C., treated with triethyl amine (8.44 mL, 60.6 mmol), and treated withdiphenylphosphoryl azide (6.53 mL, 30.3 mmol), respectively. Thereaction was allowed to warm to room temperature and stir for threehours, then concentrated in vacuo. The residue was re-dissolved in ethylacetate and washed with 1 w/w % citric acid solution. The organic wasdried over MgSO₄, filtered, and concentrated (<30° C.) to afford theintermediate azide as a yellow oil. The material was immediatelydissolved in 80 mL of tert-butanol and treated with SnCl₄ (1.65 mL of a1.0M sol'n in DCM, 1.65 mmol). The solution was heated to 80° C. for onehour to give evolution of nitrogen gas. The reaction mixture was treatedwith saturated NaHCO₃ (20 mL), and concentrated in vacuo to give a gel.The residue was partitioned between ethyl acetate and water, and theaqueous was extracted with ethyl acetate. The organic was washed withbrine, separated, dried over MgSO₄, filtered, and concentrated in vacuo.The residue was purified by chromatography (silica gel eluted with 4:1hexanes:EtOAc, R_(f)=0.20) to give the pure4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-butyric acid ethyl esteras a colorless oil (5.61 g, 60%). ¹H NMR (CDCl₃, 400 MHz) δ 7.29 (d,J=8.4 Hz, 2H), 7.23 (d, J=8.4 Hz, 2H), 4.51 (brs, 1H), 4.12 (m, 2H),3.57 (t, J=7.6 Hz, 1H), 3.09 (m, 2H), 2.25 (m, 1H), 1.93 (m, 1H), 1.43(s, 9H), 1.20 (t, J=7.2 Hz, 3H).

Step 4: The 4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-butyric acidethyl ester (5.61 g, 16.4 mmol) was dissolved in 40 mL of THF and 10 mLof water, then cooled to 0° C. The mixture was treated with lithiumhydroxide mono-hydrate (1.38 g, 32.8 mmol) to afford a yellow solution.The ice-bath was removed, and the mixture was allowed to stir overnightto room temperature. The reaction mixture was concentrated toapproximately 15 mL and diluted with water. The aqueous was washed withethyl acetate (discarded) then treated with 3M HCl solution until acidic(pH=2-3). The resulting white precipitate was extracted with ethylacetate, and the organics were combined. The organic was washed withbrine, separated, dried over MgSO₄, filtered, and concentrated in vacuoto afford the 4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-butyric acidas a colorless oil. The material was re-dissolved in a minimal amount ofwarm (60° C.) hexanes and re-concentrated to afford the pure desiredproduct as a white foam. ¹H NMR (CDCl₃, 400 MHz) δ 12.05-9.51 (brs, 1H),7.31-7.25 (m, 4H), 4.59 (brs, 1H), 3.61 (d, J=7.6 Hz, 1H), 3.14 (brs,2H), 2.28 (m, 1H), 1.92 (m, 1H), 1.43 (s, 9H).

Step 5: The 4-piperazin-1-yl-quinazoline (60 mg, 0.21 mmol) and4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-butyric acid (62 mg. 0.21mmol) were dissolved in 1.5 mL of DCM and cooled to 0° C. The solutionwas treated with PyBrop (98 mg, 0.21 mmol) and DIEA (74 μL, 0.42 mmol),respectively. The mixture was allowed to warm to room temperatureovernight, and the contents were partitioned between ethyl acetate andsaturated NH₄Cl solution. The aqueous was extracted with ethyl acetate,and the organics were combined. The organic was washed with NaHCO₃solution, separated, dried over Na₂SO₄, filtered, and concentrated invacuo. The residue was purified by chromatography (silica gel elutedwith hexanes/EtOAc gradients) to afford the pure Boc-intermediate as acolorless oil. The material was dissolved in 1.0 mL of 1,4-dioxane andtreated with 1.0 mL of 4M HCl in dioxane (2.79 mmol). The solution wasallowed to stir at room temperature overnight to completion. Theresulting suspension was diluted with diethyl ether and isolated byvacuum filtration. The pad of solid was allowed to dry under a stream ofdry nitrogen to afford the pure4-amino-2-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-onebis-hydrochloride as a white solid (32 mg, 37%). A small amount of theproduct was free-based for analytical purposes. ¹H NMR (CDCl₃, 400 MHz)δ 8.73 (s, 1H), 7.91 (d, J=8.4 Hz, 1H), 7.81 (d, J=8.4 Hz, 1H), 7.75 (t,J=7.6 Hz, 1H), 7.47 (t, J=7.6 Hz, 1H), 7.32 (d, J=8.8 Hz, 2H), 7.25 (d,J=8.8 Hz, 2H), 4.01 (dd, J=7.2 Hz, 1H), 3.94 (m, 1H), 3.81 (m, 1H), 3.71(m, 3H), 3.62 (m, 2H), 3.25 (m, 1H), 2.69 (t, J=6.8 Hz, 2H), 2.26 (m,1H), 1.84 (m, 1H), 1.40 (brs, 2H). LCMS (APCI+) m/z 410 [M+H]⁺; Rt=1.64min.

Example 62

Preparation of(3E)-4-phenyl-2-piperazin-1-yl-1-(4-quinazolin-4-yl-piperazin-1-yl)-but-3-en-1-onetrihydrochloride

(3E)-4-Phenyl-2-piperazin-1-yl-1-(4-quinazolin-4-yl-piperazin-1-yl)-but-3-en-1-onetrihydrochloride (17 mg, 47%) was prepared by the procedures describedfor the preparation of Example 1A using(3E)-2-(4-Boc-piperazinyl)-4-phenylbut-3-enoic acid. LCMS (APCI+) m/z443 [M+H]⁺. HPLC Rt 2.31 min.

Example 63

Preparation3-amino-4-phenyl-N-(1-quinazolin-4-yl-piperidin-4-yl)-butyramidedihydrochloride

Step 1: To a solution of 4-chloroquinazoline (2.0 g, 12.2 mmol) in 45 mLIPA was added Boc-4-aminopiperidine (2.56 g, 12.8 mmol) and DIEA (3.2mL, 18.2 mmol). The reaction mixture was heated to reflux and stirred 16hours, after which the reaction mixture was cooled to room temperatureand concentrated. The residue was dissolved in EtOAc and washed withwater, 1N NaOH, brine, dried (Na₂SO₄), filtered, and concentrated toprovide 4-(4-Boc-aminopiperidin-1-yl)quinazoline, which was useddirectly in the next step.

Step 2: To a solution of crude 4-(4-Boc-aminopiperidin-1-yl)quinazolinein 40 mL 1:1 dioxane:DCM was added 20 mL 4M HCl/dioxane. The resultingsuspension was stirred at room temperature for 14 hours, after which itwas concentrated to dryness. The residue was stirred in DCM and 1M NaOH,the phases were separated, and the aqueous phase was extracted with DCM.The combined organic phases were dried (Na₂SO₄), filtered andconcentrated. The residue was purified on silica gel (8:1 EtOAc:MeOH tofurnish 4-(4-aminopiperidin-1-yl)quinazoline (2.7 g, 96%) as a yellowoil. ¹H NMR (CDCl₃, 400 MHz) δ 8.73 (s, 1H), 7.92-7.85 (m, 2H),7.76-7.70 (m, 1H), 7.48-7.42 (m, 1H), 4.35-4.26 (m, 2H), 3.26-3.17 (m,2H), 3.09-2.99 (m, 1H), 2.05-1.96 (m, 2H), 1.66-1.52 (m, 2H), 1.47 (brs, 2H). LCMS (APCI+) m/z 329 [M+H]⁺. HPLC Rt 1.57 min.

Step 3: 3-Amino-4-phenyl-N-(1-quinazolin-4-yl-piperidin-4-yl)-butyramidedihydrochloride (10 mg, 33%) was prepared from4-(4-aminopiperidin-1-yl)quinazoline according to the procedure employedfor Example 1A, Step 2, using Boc-β-homophenylalanine. LCMS (APCI+) m/z390 [M+H]⁺. HPLC Rt 1.94 min.

Example 64

Preparation of3-amino-4-phenyl-N-(1-quinazolin-4-yl-piperidin-4-ylmethyl)-butyramidedihydrochloride

3-Amino-4-phenyl-N-(1-quinazolin-4-yl-piperidin-4-ylmethyl)-butyramidedihydrochloride (13 mg, 42%) was prepared by the procedures describedfor the preparation of Example 63 using 4-Boc-aminomethylpiperidine inthe S_(N)Ar step. LCMS (APCI+) m/z 404 [M+H]⁺. HPLC Rt 1.98 min.

Example 65

Preparation of3-amino-4-phenyl-1-[4-(quinazolin-4-ylamino)-piperidin-1-yl]-butan-1-onedihydrochloride

3-Amino-4-phenyl-1-[4-(quinazolin-4-ylamino)-piperidin-1-yl]-butan-1-onedihydrochloride (19 mg, 59%) was prepared by the procedures describedfor the preparation of Example 63 using 4-amino-1-Boc-piperidine in theS_(N)Ar step. LCMS (APCI+) m/z 390 [M+H]⁺. HPLC Rt 2.24 min.

Example 66

Preparation of3-amino-4-phenyl-1-[3-(quinazolin-4-ylamino)-pyrrolidin-1-yl]-butan-1-onedihydrochloride

3-Amino-4-phenyl-1-[3-(quinazolin-4-ylamino)-pyrrolidin-1-yl]-butan-1-onedihydrochloride (16 mg, 51%) was prepared by the procedures describedfor the preparation of Example 63 using 3-amino-Boc-pyrrolidine in theS_(N)Ar step. LCMS (APCI+) m/z 376 [M+H]⁺. HPLC Rt 2.17 min.

Example 67

Preparation of3-amino-4-phenyl-N-[3-(quinazolin-4-ylamino)-propyl]-butyramidedihydrochloride

3-Amino-4-phenyl-N-[3-(quinazolin-4-ylamino)-propyl]-butyramidedihydrochloride (14 mg, 46%) was prepared by the procedures describedfor the preparation of Example 63 using Boc-propylenediamine in theS_(N)Ar step. LCMS (APCI+) m/z 364 [M+H]⁺. HPLC Rt 2.14 min.

Example 68

Preparation of3-amino-4-phenyl-N-[3-(quinazolin-4-ylamino)-ethyl]-butyramidedihydrochloride

3-Amino-4-phenyl-N-[3-(quinazolin-4-ylamino)-ethyl]-butyramidedihydrochloride (11 mg, 37%) was prepared by the procedures describedfor the preparation of Example 63 using Boc-ethylenediamine in theS_(N)Ar step. LCMS (APCI+) m/z 350 [M+H]⁺. HPLC Rt 2.07 min.

Example 69

Preparation of3-Amino-4-phenyl-N-(3-quinazolin-4-yl-3-aza-bicyclo[3.1.0]hex-6-yl)-butyramidedihydrochloride

3-Amino-4-phenyl-N-(3-quinazolin-4-yl-3-aza-bicyclo[3.1.0]hex-6-yl)-butyramidedihydrochloride (19 mg, 59%, mixture of diastereomers) was prepared bythe procedures described for the preparation of Example 63 using(1α,5α,6α)-6-Boc-amino-3-azabicyclo[3.1.0]hexane (prepared according tothe literature: Tamim F. Braish et al. 1996, 1100-1102) in the S_(N)Arstep. LCMS (APCI+) m/z 388 [M+H]⁺. HPLC Rt 2.13, 2.23 min.

Example 70

Preparation of(2R)-2-amino-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: A solution containing 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (5.0g, 32.6 mmol), Boc-piperazine (15 g, 81 mmol), and DIEA (19.8 mL, 114mmol) in 130 mL IPA was stirred at 80 C for 18 hours, after which thereaction was concentrated. The crude was flashed on silica gel (20:1DCM:MeOH) to give a yellow powder, which was recrystallized fromMeOH/minimal DCM to give4-Boc-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine as a white crystallinesolid (3 crops). ¹H NMR (CDCl₃, 400 MHz) δ 11.37 (br s, 1H), 8.34 (s,1H), 7.12 (d, J=3.3 Hz, 1H), 6.49 (d, J=3.5 Hz, 1H), 4.01-3.94 (m, 4H),3.64-3.56 (m, 4H), 1.48 (s, 9H). HPLC Rt 2.05 min.

Step 2: To a solution of4-Boc-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine in 225 mL DCM was addeddropwise by addition funnel 120 mL 4M HCl/dioxane, and the resultingsuspension was stirred at room temperature 18 hours. The reactionmixture was then diluted with ether, and the solids were isolated byfiltration through a fritted funnel with nitrogen pressure, rinsed withether, and dried in vacuo to give4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride (8.44 g,94%) as a white powder. ¹H NMR (DMSO-d6, 400 MHz) δ 12.75 (1H, s), 9.63(2H, s), 8.44 (1H, s), 7.50 (1H, s), 6.95 (1H, s), 4.26-4.21 (4H, m),3.33-3.26 (4H, m). LC/MS (APCI+) m/z 204 [M+H]⁺.

Step 3: To a solution of 4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidinedihydrochloride (30 mg, 0.11 mmol), HOBt.H₂O (17 mg, 0.11 mmol), TEA (45μL, 0.33 mmol), and (D)-Boc-4-chlorophenylalanine (39 mg, 0.13 mmol) in1.6 mL DMF was added DCC (27 mg, 0.13 mmol.) The reaction mixture wasstirred at room temperature for 4 hours, after which it wasconcentrated. The residue was suspended in DCM, and the solids wereremoved by vacuum filtration through cotton plug and rinsed with DCM.The filtrate was concentrated, and the crude purified on silica gel (1:1to 1:4 DCM:EtOAc) to afford(2R)-2-Boc-amino-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one,which was used in the next step.

Step 4: To a solution of(2R)-2-Boc-amino-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onein 1 mL dioxane was added 1 mL 4M HCl/dioxane. The resulting suspensionwas stirred at room temperature overnight, after which it wasconcentrated to dryness. The solids were dissolved in minimal MeOH andthen triturated with ether. The resulting solids were isolated byfiltration through a fritted funnel with nitrogen pressure, rinsed withether, and dried in vacuo to give(2R)-2-amino-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (19 mg, 38%) as a pink powder. ¹H NMR (D₂O, 400 MHz) δ8.16 (1H, s), 7.27 (1H, s), 7.18-7.11 (4H, m), 6.68 (1H, s), 4.60-4.56(1H, m), 3.98-3.69 (4H, m), 3.61-3.52 (2H, m), 3.45-3.37 (1H, m),3.20-3.12 (1H, m), 3.00-2.91 (2H, m). LCMS (APCI+) m/z 385 [M+H]⁺. HPLCRt 1.68 min.

Example 71

Preparation of(S)-2-Aminomethyl-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: nBuLi (1.6M in hexanes, 20 mL, 32 mmol) was added to a stirredsolution of (4R,5S)-4-Methyl-5-phenyl-oxazolidin-2-one (5.2 g, 29 mmol)in THF (60 mL) at −78 C under N2. The solution was stirred at −78° C.for 10 mL and then 3-(4-Chlorophenyl)-propionyl chloride (6.0 g, 29mmol) was added and the solution allowed to warm to room temperatureover 1 hour. The solution was quenched with saturated aqueous NH4Cl,extracted into DCM (2×200 mL), dried over Na₂SO₄ and concentrated invacuo. The product was purified by column chromatography on silica (50%EtOAc/hexanes) to give(4R,5S)-3-[3-(4-Chlorophenyl)-propionyl]-4-methyl-5-phenyl-oxazolidin-2-one(4.8 g, 48%.) ¹H NMR (CDCl₃, 400 MHz) δ 7.44-7.35 (3H, m), 7.30-7.24(4H, m), 7.19 (2H, d, J 8.0 Hz), 5.64 (1H, d, J 7.4 Hz), 4.77-4.71 (1H,m), 3.34-3.17 (2H, m), 2.98 (2H, t, J 7.7 Hz), 0.88 (3H, d, J 6.7 Hz.)

Step 2: NaHMDS (1.0M, 17 mL, 17 mmol) was added to a stirred solution of(4R,5S)-3-[3-(4-Chlorophenyl)-propionyl]-4-methyl-5-phenyl-oxazolidin-2-one(4.8 g, 14 mmol) in THF (200 mL) at −78° C. under N₂. Stirred at −78° C.for 45 minutes and then Bromo-acetic acid tert-butyl ester (2.5 mL, 17mmol) was added dropwise over 10 minutes. The solution was allowed towarm to −20° C. over 4 hours and then quenched with saturated aqueousNH₄Cl. The product was extracted into EtOAc (2×300 mL), dried overNa₂SO₄, concentrated in vacuo and purified by column chromatography onsilica (20% EtOAc/hexanes) to give(3S)-3-(4-Chlorobenzyl)-4-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-yl)-4-oxo-butyricacid tert-butyl ester (5.1 g, 80%.) ¹H NMR (CDCl₃, 400 MHz) δ 7.44-7.21(9H, m), 5.45 (1H, d, J 7.3 Hz), 4.68-4.62 (1H, m), 4.51-4.42 (1H, m),3.01 (1H, dd, J 13.0 and 6.2 Hz), 2.78 (1H, dd, J 16.7 and 10.6 Hz),2.63 (1H, dd, J 13.2 and 9.0 Hz), 2.32 (1H, dd, J 6.7 and 4.3 Hz), 1.38(9H, s), 0.89 (3H, d, J 6.6 Hz.)

Step 3: A solution of(3S)-3-(4-Chlorobenzyl)-4-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-yl)-4-oxo-butyricacid tert-butyl ester (5.1 g, 11 mmol) in DCM (100 mL) was treated withTFA (50 mL) and stirred at room temperature for 1 hour. The solution wasconcentrated in vacuo, taken up into toluene and then conc. in vacuo.Placed on high vacuum for 6 hours to give(3S)-3-(4-Chlorobenzyl)-4-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-yl)-4-oxo-butyricacid (4.5 g, 100%.) ¹H NMR (CDCl₃, 400 MHz) δ 7.44-7.37 (3H, m),7.31-7.21 (6H, m), 5.48 (1H, d, J 6.9 Hz), 4.70-4.63 (1H, m), 4.50-4.42(1H, m), 3.05 (1H, dd, J 13.3 and 6.2 Hz), 2.89 (1H, dd, J 17.7 and 10.6Hz), 2.63 (1H, dd, J 13.2 and 9.0 Hz), 2.43 (1H, dd, J 17.6 and 4.4 Hz),0.86 (3H, d, J 6.6 Hz.)

Step 4: NEt3 (700 uL, 5.0 mmol) was added to a stirred solution of(3S)-3-(4-Chlorobenzyl)-4-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-yl)-4-oxo-butyricacid (1.0 g, 2.5 mmol) in PhMe (50 mL) at 0° C. under N₂. This wasfollowed by the addition of the diphenylphosphoryl azide (650 μL, 3.0mmol.) The solution was stirred at 0° C. for 15 minutes and then stirredat room temperature overnight. The solution was washed with 1% citricacid, extracted into EtOAc and concentrated in vacuo. Taken up intotBuOH (50 mL), SnCl4 (1.0M in DCM, 0.1 mL) added and stirred and heatedat 85° C. for 5 hours. Cooled to room temperature and quenched withsaturated aqueous bicarbonate. Stirred at RT for 10 minutes and thenconcentrated in vacuo. The product was taken up into water (100 mL) andextracted into EtOAc (2×200 mL.) Dried over Na₂SO₄ and concentrated invacuo. The product was purified by column chromatography on silica (10%EtOAc/hexanes) and then on the Biotage to give[(2S)-2-(4-Chlorobenzyl)-3-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-yl)-3-oxo-propyl]-carbamicacid tert-butyl ester (300 mg, 25%.) LCMS (APCI+) m/z 373 [M-Boc+H]⁺;Rt: 3.92 min.

Step 5: To a solution of[(2S)-2-(4-Chlorobenzyl)-3-((4R,5S)-4-methyl-2-oxo-5-phenyl-oxazolidin-3-yl)-3-oxo-propyl]-carbamicacid tert-butyl ester (300 mg, 0.63 mmol) in THF/H₂O (30/10 mL) at 0° C.was added LiOH (80 mg, 1.9 mmol) and H₂O₂ (30% by volume, 3.0 mL, 0.63mmol) and stirred at 0° C. for 30 minutes. Then Na₂SO₃ (saturatedsolution, 10 mL) was added slowly & cautiously. Diluted with EtOAc (100mL) and extracted into water (2×100 mL.) The aq. layer was acidified (1NHCl) and extracted into EtOAc (3×100 mL.) Dried over Na₂SO₄ andconcentrated in vacuo to give(S)-2-(tert-Butoxycarbonylamino-methyl)-3-(4-chlorophenyl)-propionicacid (150 mg, 75%.) LCMS (APCI—) m/z 322 [M-Boc+H]⁻; Rt: 2.23 min.

Step 6: NEt₃ (150 μL, 1.1 mmol) was added to a stirred suspension of4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride (100 mg,0.36 mmol),(S)-2-(tert-Butoxycarbonylamino-methyl)-3-(4-chlorophenyl)-propionicacid (130 mg, 0.40 mmol), EDCI (83 mg, 0.44 mmol) and HOBt (59 mg, 0.44mmol) in DMF (15 mL) at RT. Stirred at RT overnight. Poured into EtOAc(100 mL), washed with water (100 mL), 1N NaOH (50 mL), dried overNa2SO4, concentrated in vacuo, purified by column chromatography onsilica (100% EtOAc) to give(S)-{2-(4-Chlorobenzyl)-3-oxo-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propyl}-carbamicacid tert-butyl ester. LCMS (APCI+) m/z 499 [M+H]⁺; Rt: 2.70 minutes.This was taken up into DCM (50 mL) and stirred with TFA (5 mL)overnight. Poured into EtOAc (100 mL) and washed with 1N NaOH (2×100mL), dried over Na₂SO₄ and concentrated in vacuo. Formed HCl salt(Et₂O.HCl, 2M in diethyl ether) to give(S)-2-Aminomethyl-3-(4-chlorophenyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (100 mg, 59%.) LCMS (APCI+) m/z 399 [M+H]⁺; Rt: 1.82min. [Free base: ¹H NMR (CDCl₃, 400 MHz) δ 9.97 (1H, br.s), 8.32 (1H,s), 7.24 (2H, d, J 7.4 Hz), 7.13 (2H, d, J 7.4 Hz), 7.09 (1H, d, J 2.4Hz), 6.45 (1H, d, J 2.3 Hz), 4.00-3.68 (5H, m), 3.60-3.53 (1H, m),3.46-3.40 (1H, m), 3.33-3.27 (1H, m), 3.16-3.05 (2H, m), 2.92-2.83 (2H,m), 2.77 (1H, dd, J 13.3 and 5.5 Hz.)]

Example 72

Preparation of(R)-2-Amino-1-[4-(3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazin-1-yl]-3-(4-chlorophenyl)-propan-1-one,dihydrochloride

Step 1: The 4-hydroxypyrazolopyrimidine (2.5 g, 18 mmol) was dissolvedin POCl₃ (34 mL, 0.37 mol) and N,N-dimethyl aniline (4.7 mL, 37 mmol.)This mixture was heated to reflux (120° C.) for 1.5 hours to afford adark red solution. The mixture was concentrated to a viscous oil andcooled to 0° C. in an ice bath. The oil was poured into a mixture ofice-water and was stirred for 5 minutes. The acidic melt was extractedwith ether (4×100 mL), and the organics were combined. The organic waswashed with cold water, then cold half saturated NaHCO3 solution, thenbrine, separated, dried over MgSO₄, filtered, and concentrated in vacuoto afford 4-Chloro-1H-pyrazolo[3,4-d]pyrimidine (1.1 g, 39%) as a lightyellow powder. ¹H NMR (DMSO-d6, 400 MHz) δ 8.79 (1H, s), 8.41 (1H, s.)

Step 2: To a suspension of 4-Chloro-1H-pyrazolo[3,4-d]pyrimidine (1.1 g,7.1 mmol) in CHCl₃ (50 mL) was added NBS (1.49 g, 8.4 mmol.) The mixturewas stirred at room temperature for 5 hours, cooled to 0 C and thesolids were isolated by vacuum filtration, rinsed with cold CHCl₃, andair dried. The solid was purified by column chromatography on silica(50% EtOAc/hexanes) to give3-Bromo-4-chloro-1H-pyrazolo[3,4-d]pyrimidine (1.3, 77%.)

Step 3: To a solution of 3-Bromo-4-chloro-1H-pyrazolo[3,4-d]pyrimidine(1.3 g, 5.5 mmol) in DMF (42 mL) at 0° C. was added NaH (180 mg, 7.7mmol) in portions. The reaction mixture was stirred at 0° C. for 5minutes, then stirred at room temperature for 1.5 hours, after which itwas cooled back to 0° C. Neat PhSO₂Cl (0.7 mL, 5.6 mmol) was added andthe reaction mixture was warmed to room temperature and stirredovernight. The reaction was quenched with saturated aqueous NH₄Cl anddiluted further with H₂O. The resulting precipitate was isolated byvacuum filtration to give1-Benzenesulfonyl-3-bromo-4-chloro-1H-pyrazolo[3,4-d]pyrimidine (1.8 g,88%.) ¹H NMR (DMSO-d6, 400 MHz) δ 9.11 (1H, s), 8.11 (2H, d, J 8.1 Hz),7.84 (1H, t, J 7.5 Hz), 7.70 (2H, t, J 8.0 Hz.)

Step 4: A solution of1-Benzenesulfonyl-3-bromo-4-chloro-1H-pyrazolo[3,4-d]pyrimidine (1.8 g,4.8 mmol), Boc-piperazine (1.4 g, 7.2 mmol) and DIPEA (2.1 mL, 12 mmol)in IPA (40 mL) was stirred and heated at reflux overnight. The reactionmixture was cooled to −10° C., the solids isolated by vacuum filtration,rinsed with cold IPA and dried further on high vacuum line to give4-(1-Benzenesulfonyl-3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (1.7 g, 67%) as a white powder. LCMS (APCI+) m/z523 and 525 [M+H]⁺; Rt: 3.57 min. ¹H NMR (CDCl₃, 400 MHz) δ 8.53 (1H,s), 8.23 (2H, d, J 8.5 Hz), 7.65 (1H, t, J 6.9 Hz), 7.54 (2H, t, J 7.8Hz), 3.82-3.79 (4H, m), 3.60-3.57 (4H, m), 1.48 (9H, s.)

Step 5: Anhydrous HCl (4N in dioxane, 10 mL) was added to a stirredsolution of1-Benzenesulfonyl-3-bromo-4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidine(210 mg, 0.40 mmol) in MeOH (20 mL) and stirred at room temperatureovernight. The suspension was concentrated in vacuo to give1-Benzenesulfonyl-3-bromo-4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidinedihydrochloride (200 mg, 100%.) LCMS (APCI+) m/z 423 and 425 [M+H]⁺; Rt:1.98 min.

Step 6: DIPEA (84 ul, 0.48 mmol) was added to a suspension of1-Benzenesulfonyl-3-bromo-4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidinedihydrochloride (40 mg, 0.081 mmol) and(D)-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid (27 mg,0.089 mmol) in DCM (10 mL) at room temperature. Then, HBTU (34 mg, 0.089mmol) was added and the reaction stirred at room temperature overnight.MeOH (5 mL) and 3M LiOH (3 mL) were added and the mixture stirred andheated at 50° C. for 2 hours. The mixture was diluted with saturatedaqueous NaHCO₃ (10 mL), extracted into DCM, dried over Na₂SO₄ andconcentrated in vacuo. The crude mixture was purified by columnchromatography on silica (50% EtOAc/hexanes) to give(R)-[2-[4-(3-Bromo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazin-1-yl]-1-(4-chlorobenzyl)-2-oxo-ethyl]-carbamicacid tert-butyl ester. LCMS (APCI+) m/z 564 and 566 [M+H]⁺; Rt: 3.04min. This was taken up into MeOH (10 mL) and treated with anhydrous HCl(4M in dioxane, 20 mL.) The solution was stirred at room temperatureovernight and concentrated in vacuo to give(R)-2-Amino-1-[4-(3-bromo-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazin-1-yl]-3-(4-chlorophenyl)-propan-1-one,dihydrochloride (13 mg, 30%.) LCMS (APCI+) m/z 464 and 466 [M+H]⁺; Rt:2.00 min.

Example 73

Preparation of(S)-2-Aminomethyl-3-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a suspension of 4-Chloro-7H-pyrrolo[2,3-d]pyrimidine (2.5 g,16 mmol) in CDCl₃ (65 mL) was added NBS (2.9 g, 16 mmol) and thereaction mixture stirred and heated at reflux for 2.5 hours. The mixturewas cooled to room temperature, the solids isolated by vacuumfiltration, rinsed with cold CHCl₃ and air dried to give5-Bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (3.0 g, 79%.) LCMS (APCI+)m/z 232 and 234 [M+H]⁺; Rt: 2.32 min. ¹H NMR (DMSO-d6, 400 MHz) δ 12.98(1H, br. s), 8.63 (1H, s), 7.96 (1H, s.)

Step 2: To a solution of 5-Bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine(3.0 g, 13 mmol) in DMF (40 mL) at 0° C. was added NaH (60% w/w inmineral oil, 720 mg, 18.1 mmol) and the mixture stirred at 0° C. underN₂ for 30 minutes. Then PhSO₂Cl (1.7 g, 13 mmol) was added and thereaction stirred at room temperature for 2 hours, after which H₂O (200mL) was added, causing precipitation. The precipitate was collected byfiltration and dried under vacuum to give7-Benzenesulfonyl-5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (4.9 g,100%.) ¹H NMR (DMSO-d6, 400 MHz) δ 8.85 (1H, s), 8.45 (1H, s), 8.19 (2H,d, J 8.7 Hz), 7.81 (1H, t, J 7.4 Hz), 7.70 (2H, t, J 7.8 Hz.)

Step 3: A suspension of7-Benzenesulfonyl-5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (4.9 g,13 mmol), piperazine-1-carboxylic acid tert-butyl ester (3.7 g, 20mmol), and DIPEA (5.7 mL, 33 mmol) in IPA (30 mL) was stirred and heatedat reflux for 6 hours. The mixture was cooled to −10° C., the solidscollected by vacuum filtration, rinsed with cold IPA and dried undervacuum to give4-(7-Benzenesulfonyl-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (5.9 g, 86%.) LCMS (APCI+) m/z 522 and 524 [M+H]⁺;Rt: 3.92 min. ¹H NMR (DMSO-d6, 400 MHz) δ 8.48 (1H, s), 8.21 (2H, d, J8.2 Hz), 7.66-7.62 (2H, m), 7.54 (2H, t, J 7.8 Hz), 3.62-3.55 (8H, m),1.48 (9H, s.)

Step 4: MeZnCl (2.0M in THF, 720 uL, 1.4 mmol) was added to a stirredsolution4-(7-Benzenesulfonyl-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (250 mg, 0.48 mmol) andtetrakis(triphenylphosphine)palladium(0) (140 mg, 0.12 mmol) in THF (10mL) at room temperature under N₂. The solution was stirred and heated atreflux for 2 hours, cooled to room temperature, quenched with saturatedaqueous NH₄Cl, extracted into EtOAc (2×100 mL), dried over Na₂SO₄ andconcentrated in vacuo. The product was purified on a Biotage (silica,40% EtOAc/hexanes) to give4-(7-Benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (210 mg, 94%.) LCMS (APCI+) m/z 458 [M+H]⁺; Rt:3.73. ¹H NMR (CDCl₃, 400 MHz) δ 8.49 (1H, s), 8.18 (2H, d, J 8.2 Hz),7.59 (1H, t, J 6.9 Hz), 7.50 (2H, t, J 7.8 Hz), 7.34 (1H, s), 3.59-3.54(4H, m), 3.48-3.44 (4H, m), 2.35 (3H, s), 1.48 (9H, s.)

Step 5: A solution of4-(7-Benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (4.0 g, 13 mmol) in DCM (100 mL) was treated withanhydrous HCl (4M in dioxane, 100 mL) and stirred at room temperatureovernight. The suspension was concentrated in vacuo to give7-Benzenesulfonyl-5-methyl-4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidinedihydrochloride (5.4 g, 100%.) LCMS (APCI+) m/z 358 [M+H]⁺; Rt: 1.79.

Step 6: DIPEA (120 L, 0.70 mmol) was added to a suspension of7-Benzenesulfonyl-5-methyl-4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidinedihydrochloride (50 mg, 0.12 mmol) and(S)-2-(tert-Butoxycarbonylamino-methyl)-3-(4-chlorophenyl)-propionicacid (40 mg, 0.13 mmol) in DCM (10 mL) at room temperature. Then, HBTU(48 mg, 0.13 mmol) was added and the reaction stirred at roomtemperature overnight. MeOH (5 mL) and 3M LiOH (1.2 mL) were added andthe mixture stirred and heated at 50° C. for 2 hours. The mixture wasdiluted with saturated aqueous NaHCO₃ (10 mL), extracted into DCM, driedover Na₂SO₄ and concentrated in vacuo. The crude mixture was purified bycolumn chromatography on silica (50% EtOAc/hexanes) to give(S)-{2-(4-Chlorobenzyl)-3-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-3-oxo-propyl}-carbamicacid tert-butyl ester. LCMS (APCI+) m/z 253 [M+H]⁺; Rt: 2.85 min. Thiswas taken up into MeOH (10 mL) and treated with anhydrous HCl (4M indioxane, 20 mL.) The solution was stirred at room temperature overnightand concentrated in vacuo to give(S)-2-Aminomethyl-3-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (21 mg, 37%.) LCMS (APCI+) m/z 413 [M+H]⁺; Rt: 1.82 min.

Example 73

The Preparation of(2S)-2-Aminomethyl-3-(4-chlorophenyl)-1-[4-(5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a solution of4-(7-Benzenesulfonyl-5-bromo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (150 mg, 0.29 mmol) in 4 mL DME (degassed withnitrogen prior to use) was added 0.94M aqueous Na₂CO₃ (0.61 mL, 0.57mmol) and Pd(PPh₃)₄ (66 mg, 0.057 mmol). The reaction mixture wasstirred 5 minutes, then 2-thiophene boronic acid (55 mg, 0.43 mmol) wasadded. The reaction mixture heated to reflux and stirred 16 hours, afterwhich it was cooled to room temperature and DME was removed by rotaryevaporation. The reaction mixture was diluted with H₂O and extractedwith DCM, and the combined extracts were dried (Na₂SO₄), filtered, andconcentrated. The crude was purified on silica gel (8:1 to 4:1hexanes:EtOAc) to furnish4-(7-Benzenesulfonyl-5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (120 mg, 79%) as a beige powder. LCMS (APCI+) m/z526 [M+H]⁺; Rt: 3.25 min.

Step 2: To a solution of4-(7-Benzenesulfonyl-5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (120 mg, 0.23 mmol) in 2 mL dioxane was added 1.5mL 4M HCl/dioxane. The reaction mixture was stirred at room temperature6 hours, after which it was diluted with ether, and the solids wereisolated by filtration through a fritted funnel with nitrogen pressure,rinsed with ether, and dried in vacuo to give7-Benzenesulfonyl-4-piperazin-1-yl-5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidinedihydrochloride (110 mg, 95%) as a yellow powder. LCMS (APCI+) m/z 426[M+H]⁺; Rt: 1.95 min.

Step 3: To a solution of7-Benzenesulfonyl-4-piperazin-1-yl-5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidinedihydrochloride (50 mg, 0.10 mmol), DIEA (0.10 mL, 0.60 mmol), and(2S)-2-(Boc-aminomethyl)-3-(4-chlorophenyl)-propionic acid (38 mg, 0.12mmol) in 2 mL DCM was added HBTU (44 mg, 0.12 mmol). The reactionmixture was stirred at room temperature 2 hours, after which 2 mL MeOHand 0.5 mL 3M LiOH were added. The reaction mixture was heated to 35° C.and stirred 2 hours, after which saturated NaHCO₃ was added, and themixture was extracted with DCM. The combined extracts were dried(Na₂SO₄), filtered, and concentrated. The crude was purified on silicagel (flushed with 2:1 DCM:EtOAc, then gradient to 1:4 DCM:EtOAc) to give(2S)-2-Boc-aminomethyl-3-(4-chlorophenyl)-1-[4-(5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pip-erazin-1-yl]-propan-1-one,which was used in the next step.

Step 4: To a solution of(2S)-2-Boc-aminomethyl-3-(4-chlorophenyl)-1-[4-(5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-pip-erazin-1-yl]-propan-1-onein 1.5 mL dioxane was added 1.5 mL 4M HCl/dioxane. The resultingsuspension was stirred at room temperature 16 hours, after which it wasconcentrated to dryness. The solids were dissolved in minimal MeOH, andthe product was triturated by the addition of ether. The solids wereisolated by filtration through a fritted funnel with nitrogen pressure,rinsed with ether, and dried in vacuo to give(2S)-2-aminomethyl-3-(4-chlorophenyl)-1-[4-(5-thiophen-2-yl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (37 mg, 67%) a pale yellow powder. ¹H NMR (D₂O, 400 MHz)δ 8.18 (1H, s), 7.41 (1H, d, J 5.4 Hz), 7.30 (1H, s), 7.19 (2H, d, J 8.6Hz), 7.06 (1H, dd, J 5.0 and 3.5 Hz), 7.01 (2H, d, J 8.0 Hz), 6.88 (1H,d, J 3.5 Hz), 3.47-3.14 (5H, m), 3.08-3.00 (3H, m), 2.86-2.77 (3H, m),2.59 (1H, t, J 12.0 Hz), 2.13-2.06 (1H, m). LCMS (APCI+) m/z 481 [M+H]⁺;Rt: 1.87 min.

Example 75

Preparation of2-Amino-3-(4-chloro-2-methyl-phenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one,dihydrochloride

Step 1: A mixture containing 4-Chloro-2-methyl-benzoic acid (4 g, 23mmol) and LiAlH4 (890 mg, 23.5 mmol) in 250 mL of THF under a nitrogenatmosphere was allowed to stir at room temperature for 2 hours. Thereaction was quenched with sodium sulfate decahydrate. The mixture wasfiltered through a pad of Celite and the filter cake washed with THF.The filtrate was concentrated under reduced pressure. Purification ofthe residue via biotage eluting with 30% ethyl acetate/hexanes gave(4-Chloro-2-methyl-phenyl)-methanol (3.70 g, 100%) as a colorless oil.¹H NMR (CDCl₃, 400 MHz) δ 7.30-7.25 (1H, m), 7.18-7.14 (2H, m), 4.66(2H, d, J 5.8 Hz), 2.32 (3H, s.)

Step 2: A solution containing gave (4-Chloro-2-methyl-phenyl)-methanol(2 g, 13 mmol) and PBr3 (1.3 mL, 14 mmol) in 150 mL of diethyl ether wasallowed to stir at room temperature overnight. The reaction was dilutedwith ether and washed with water. The organic phase was dried overmagnesium sulfate. Filtration, removal of solvent and purification ofthe residue via biotage eluting with 20% ethyl acetate/hexanes gave1-Bromomethyl-4-chloro-2-methyl-benzene (1.89 g, 67%) as a colorlessoil. ¹H NMR (CDCl₃, 400 MHz) δ 7.26-7.11 (3H, m), 4.67 (2H, s), 2.39(3H, s.)

Step 3: To a solution containing (Benzhydrylidene-amino)-acetic acidethyl ester (2.3 g, 8.6 mmol) in 50 mL of DMSO under a nitrogenatmosphere was added potassium t-butoxide (1.2 g, 11 mmol) Afterstirring for 20 minutes, 1-Bromomethyl-4-chloro-2-methyl-benzene (1.89g, 8.6 mmol) was added and the reaction allowed to stir at roomtemperature overnight. The reaction was diluted with ethyl acetate andwashed with brine. The organic phase was dried over magnesium sulfate.Filtration, removal of solvent and purification of the residue viabiotage eluting with 20% ethyl acetate/hexanes gave2-(Benzhydrylidene-amino)-3-(4-chloro-2-methyl-phenyl)-propionic acidethyl ester (1.75 g, 50%) as a yellow oil. LCMS (APCI+) m/z 406 [M+H]⁺;Rt: 4.16 min.

Step 4: A mixture containing2-(Benzhydrylidene-amino)-3-(4-chloro-2-methyl-phenyl)-propionic acidethyl ester (1.7 g, 4.2 mmol) and 90 mL of 3N HCl was heated at 75 Covernight. The reaction was cooled to room temperature and washed withethyl acetate. The aqueous phase was concentrated under reduced pressureto afford 2-Amino-3-(4-chloro-2-methyl-phenyl)-propionic acid (640 mg,72%) as white solid. LCMS (APCI+) m/z 214 [M+H]⁺; Rt: 1.83 min. ¹H NMR(D₂O, 400 MHz) δ 7.18 (1H, s), 7.11 (1H, d, J 8.7 Hz), 7.05 (1H, d, J8.3 Hz), 4.05-4.00 (1H, m), 3.23 (1H, dd, J 14.3 and 6.1 Hz), 2.98 (1H,dd, J 14.4 and 8.5 Hz), 2.19 (3H, s.)

Step 5: To a solution containing2-Amino-3-(4-chloro-2-methyl-phenyl)-propionic acid (640 mg, 3.0 mmol),25 mL of dioxane and 9 mL of 1N sodium hydroxide was added boc anhydride(0.73 g, 3.3 mmol.) The reaction was allowed to stir at room temperaturefor 3 hours. The reaction was diluted with water and washed with DCM.The aqueous phase was acidified with 1N HCl and extracted with ethylacetate. The organic phase was dried over magnesium sulfate. Filtrationand removal of solvent gave2-tert-Butoxycarbonylamino-3-(4-chloro-2-methylphenyl)-propionic acid(640 mg, 80%) as a white solid. ¹H NMR (CDCl₃, 400 MHz) δ 7.13 (1H, s),7.07 (2H, s), 5.14-5.08 (1H, m), 4.55-4.48 (1H, m), 3.24-3.16 (1H, m),2.96-2.88 (1H, m), 2.33 (3H, s), 1.38 (9H, s.)

Step 6: To a solution containing2-tert-Butoxycarbonylamino-3-(4-chloro-2-methyl-phenyl)-propionic acid(200 mg, 0.64 mmol) in 30 mL of DMF was added HOBT (0.12 g, 0.76 mmol),EDCI (0.15 g, 0.76 mmol) and NMM (0.19 g, 1.9 mmol) under a nitrogenatmosphere. After stirring for 10 minutes, 4-piperazin-1-yl-quinazoline(200 mg, 0.93 mmol) was added and stirring continued overnight. Thereaction was diluted with ethyl acetate and washed with water. Theorganic phase was dried over magnesium sulfate. Filtration, removal ofsolvent and purification of the residue via biotage eluting with 10%MeOH/DCM gave[1-(4-Chloro-2-methylbenzyl)-2-oxo-2-(4-quinazolin-4-yl-piperazin-1-yl)-ethyl]-carbamicacid tert-butyl ester (0.31 g, 95%.) LCMS (APCI+) m/z 510 [M+H]⁺; Rt:2.54 min.

Step 7: To a solution containing[1-(4-Chloro-2-methylbenzyl)-2-oxo-2-(4-quinazolin-4-yl-piperazin-1-yl)-ethyl]-carbamicacid tert-butyl ester (0.30 g, 0.59 mmol) in 30 mL of DCM under anitrogen atmosphere was added TFA (1.4 mL.) After stirring at roomtemperature overnight, the reaction was concentrated under reducedpressure. The residue was dissolved in DCM and 2N HCl in ether added.The solids were filtered and dried to afford2-Amino-3-(4-chloro-2-methyl-phenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-onedihydrochloride (0.217 g) as an off-white solid. LCMS (APCI+) m/z 410[M+H]⁺; Rt: 1.87 min. ¹H NMR (D2O, 400 MHz) δ 8.48 (1H, s), 7.91-7.86(2H, m), 7.66-7.59 (2H, m), 7.08-7.02 (2H, m), 4.60-4.54 (1H, m),4.20-4.12 (1H, m), 3.86-3.72 (3H, m), 3.61-3.38 (2H, m), 3.28-3.16 (2H,m), 3.01-2.94 (1H, m), 2.76-2.68 (1H, m), 2.19 (3H, s.)

Example 76

Preparation of4-{4-[3-amino-2-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-1,3-dihydropyrrolo[2,3-b]pyrimidin-2-onedihydrochloride

Step 1: A solution of 4-chloro-5,7-dihydropyrrolo[2,3-d]pyrimidin-6-one(prepared according to the literature: Li Sun et al. Bioorg. and Med.Chem. Lett. 2002, 12, 2153-2157; 690 mg, 3.7 mmol), Boc-piperazine (630mg, 3.7 mmol), and DIEA (0.96 mL, 5.5 mmol) in 20 mL IPA was heated toreflux and stirred 14 hours, after which the reaction mixture wasconcentrated. The crude was purified on silica gel (1:2 DCM:EtOAc to 1:4DCM:EtOAc gradient) to give4-Boc-piperazin-1-yl-5,7-dihydropyrrolo[2,3-d]pyrimidin-6-one, which wasused in the next step.

Step 2: To a solution of4-Boc-piperazin-1-yl-5,7-dihydropyrrolo[2,3-d]pyrimidin-6-one in 25 mLdioxane, was added 15 mL 4M HCl/dioxane. The resulting suspension wasstirred at room temperature 15 hours, after which it was diluted withether. The solids were isolated by filtration through a fritted funnelwith nitrogen pressure, rinsed with ether, and dried in vacuo to furnish4-piperazin-1-yl-5,7-dihydropyrrolo[2,3-d]pyrimidin-6-onedihydrochloride (350 mg, 100%) as a red powder. ¹H NMR (DMSO-d6, 400MHz) δ 11.12 (1H, s), 9.26 (2H, br. s), 8.25 (1H, s), 3.89-3.84 (4H, m),3.77 (2H, s), 3.16-3.09 (4H, m). LCMS (APCI+) m/z 220 [M+H]⁺; Rt: 0.68min.

Step 3: To a solution of give4-piperazin-1-yl-5,7-dihydropyrrolo[2,3-d]pyrimidin-6-onedihydrochloride (40 mg, 0.14 mmol), HOBt.H₂O (21 mg, 0.14 mmol), TEA (57μL, 0.41 mmol), and 3-Boc-amino-2-(4-chlorophenyl)-propionic acid(prepared from 4-chlorophenylacetic acid methyl ester using theprocedures described for the preparation of A109; 49 mg, 0.16 mmol) in1.2 mL 5:1 DCM:THF was added DCC (34 mg, 0.16 mmol). The reactionmixture was stirred at room temperature 4 hours, after which it wasconcentrated. The residue was suspended in DCM, and solids were removedby vacuum filtration through a cotton plug and rinsed with DCM. Thefiltrate was concentrated, and the crude was purified on silica gel (1:1to 1:5 DCM:EtOAc gradient) to give4-{4-[3-Boc-amino-2-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-1,3-dihydropyrrolo[2,3-b]pyrimidin-2-one,which was used in the next step. LCMS (APCI+) m/z 401 [M-Boc+H]⁺; Rt:2.16 min.

Step 4: To a solution of4-{4-[3-Boc-amino-2-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-1,3-dihydropyrrolo[2,3-b]pyrimidin-2-onein 1.2 mL dioxane was added 1.2 mL 4M HCl/dioxane. The resultingsuspension was stirred at room temperature 15 hours, after which it wasconcentrated to dryness. The solids were dissolved in minimal MeOH, andthe product was triturated by the addition of ether. The resultingsolids were isolated by filtration through a fritted funnel withnitrogen pressure, rinsed with ether, and dried in vacuo to give4-{4-[3-amino-2-(4-chlorophenyl)-propionyl]-piperazin-1-yl}-1,3-dihydropyrrolo[2,3-b]pyridin-2-onedihydrochloride (28 mg, 43%) as a dark pink powder. ¹H NMR (D₂O, 400MHz) δ 8.10 (1H, s), 7.31-7.14 (4H, m), 4.28-4.22 (1H, m), 3.92-3.82(1H, m), 3.73-2.94 (11H, m). LCMS (APCI+) m/z 401 [M+H]⁺; Rt: 1.98 min.

Example 77

Preparation of4-Amino-2-(4-chlorophenyl)-2-fluoro-1-(4-quinazolin-4-yl-piperazin-1-l)-butan-1-onedihydrochloride

Step 1: A solution of 4-chloromandelic acid (12.3 g, 65.9 mmol) intoluene (50 mL), EtOH (16 mL), and concentrated H₂SO₄ (0.1 mL) wasrefluxed for 12 hours while removing water using a Dean-Stark trap. Themixture was concentrated in vacuo, diluted with DCM, and washed withdilute aqueous NaHCO₃. The separated DCM layer was dried (MgSO₄),filtered, and concentrated in vacuo to give(4-chlorophenyl)-hydroxy-acetic acid ethyl ester as a colorless oil(10.0 g) that crystallized upon standing.

Step 2: (4-Chlorophenyl)-hydroxy-acetic acid ethyl ester (10.0 g, 46.6mmol) in DCM (35 mL) was cannulated into a solution of[bis(2-methoxyethyl)amino]sulfur trifluoride (9.45 mL, 51.3 mmol) in DCM(35 mL) cooled at −78° C. After being stirred for 12 hours and allowedto warm to ambient temperature, the mixture was poured into saturatedaqueous NaHCO₃. The mixture was extracted with DCM and the organicextracts were dried (MgSO₄), filtered, and concentrated in vacuo. Thecrude material was chromatographed (SiO₂) using DCM as eluent to give(4-chlorophenyl)-fluoroacetic acid ethyl ester as a colorless oil (7.0g).

Step 3: Potassium tert-butoxide (155 mg, 1.38 mmol) was added to asolution of (4-chlorophenyl)-fluoroacetic acid ethyl ester (3.00 g, 13.8mmol) in THF (25 mL) at 0° C. to give an orange-red color. After 15minutes, the mixture was cooled to −78° C. and t-butyl acrylate (2.23mL, 15.2 mmol) was added neat. After being stirred and allowed to warmto ambient temperature for 12 hours, the mixture was quenched withsaturated NH₄Cl, concentrated in vacuo, diluted with H₂O, and extractedwith DCM. The DCM extracts were dried (MgSO₄), filtered, andconcentrated in vacuo. The crude material was chromatographed (SiO₂)using DCM as eluent to give a colorless oil (1.20 g). A solution of theoil in DCM (6 mL) and TFA (4 mL) was stirred overnight. The mixture wasdiluted with toluene (40 mL) and concentrated in vacuo. The crudeproduct was dissolved in dilute aqueous NaHCO₃ and extracted with DCM(twice, discarded). The aqueous layer was acidified to pH 1.0 with 1.0 NHCl and extracted with DCM (twice). The organic extracts were dried(Na₂SO₄), filtered, and concentrated in vacuo to give2-(4-chlorophenyl)-2-fluoro-pentanedioic acid 1-ethyl ester as an oil(1.0 g).

Step 4: Triethylamine (0.53 mL, 3.81 mmol) was added to a solution of2-(4-chlorophenyl)-2-fluoro-pentanedioic acid 1-ethyl ester (1.00 g,3.46 mmol) in t-BuOH (20 mL) followed by the addition ofdiphenylphosphoryl azide (0.82 mL, 3.81 mmol). The mixture was heated at95° C. for 3 hours, concentrated in vacuo, and partitioned betweendilute aqueous NaHCO₃ and DCM. The separated DCM layer was dried(Na₂SO₄), filtered, and concentrated in vacuo. The crude product waschromatographed (SiO₂) using DCM as eluent to give4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-2-fluoro-butyric acidethyl ester (600 mg). ¹H NMR (CDCl₃, 400 MHz) δ 7.42 (m, 2H), 7.35 (m,2H), 5.25 (m, 0.4H), 4.61 (m, 0.6H), 4.19 (m, 2H), 3.35 (m, 0.6H), 3.23(m, 1.4H), 2.59 (m, 1H), 2.34 (m, 1H), 1.40 (m, 9H), 1.24 (m, 3H). LCMS(APCI+) m/z 260 [M+H]⁺ (loss of Boc group).

Step 5: Lithium hydroxide monohydrate (0.27 g, 6.45 mmol) in H₂O (5 mL)was added to a solution of4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-2-fluoro-butyric acidethyl ester (580 mg, 1.61 mmol) in THF (5 mL) and MeOH (5 mL). Afterbeing stirred for 12 hours, the mixture was concentrated in vacuo,diluted with H₂O, and extracted with DCM (3 times, discarded). Theaqueous phase was then acidified to pH 1 and extracted with DCM (2times). The organic extracts were dried (Na₂SO₄), filtered, andconcentrated in vacuo to give4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-2-fluoro-butyric acid (400mg). The material was used in the following step without furtherpurification.

Step 6: PyBrop (562 mg, 1.21 mmol) was added in a single portion to asolution of4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-2-fluoro-butyric acid (400mg, 1.21 mmol) and 4-piperazin-1-yl-quinazoline dihydrochloride (346 mg,1.21 mmol) in DCM (12 mL) cooled in an ice bath. DIEA (0.84 mL, 4.82mmol) was added and the mixture was allowed to warm to ambienttemperature and stirred for 12 hours. The mixture was diluted with DCMand washed with 0.1 N HCl. The separated DCM layer was dried (Na₂SO₄),filtered, and concentrated in vacuo. The crude material waschromatographed (SiO₂) using 2% MeOH/DCM followed by 5% MeOH/DCM aseluent to give[3-(4-chlorophenyl)-3-fluoro-4-oxo-4-(4-quinazolin-4-yl-piperazin-1-yl)-butyl]-carbamicacid tert-butyl ester (230 mg). LCMS (APCI+) m/z 528, 530 [M+H]⁺.

Step 7: A solution of[3-(4-chlorophenyl)-3-fluoro-4-oxo-4-(4-quinazolin-4-yl-piperazin-1-yl)-butyl]-carbamicacid tert-butyl ester (226 mg, 0.43 mmol) in DCM (2 mL) and 2.0 M HCl inEt₂O (1 mL) was stirred for 12 hours. The mixture was concentrated invacuo and chromatographed (SiO₂) using 10% MeOH/DCM followed by 10% (7 NNH₃ in MeOH)/DCM as eluent. The purified material was dissolved in MeOHfollowed by the addition of 2.0 N HCl in Et₂O, and then concentrated invacuo. The resulting glass was heated at reflux in isopropyl alcohol andconcentrated in vacuo. The resulting solid was suspended in acetonitrileand concentrated in vacuo (repeated twice) to give4-amino-2-(4-chlorophenyl)-2-fluoro-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-onedihydrochloride as a white powder. ¹H NMR (DMSO-d₆, 400 MHz) δ 8.86 (s,1H), 8.18 (m, 4H), 8.01 (m, 2H), 7.69 (m, 1H), 7.58 (d, J=8.4 Hz, 2H),7.40 (d, J=8.6 Hz, 2H), 4.23 (m, 2H), 4.09 (m, 1H), 3.80 (m, 4H), 3.47(m, 1H), 2.86 (m, 1H), 2.67 (m, 2H), 2.41 (m, 1H). LCMS (APCI+) m/z 428,430 [M+H]⁺.

Example 78

Preparation of3-Amino-2-(4-bromo-2-fluoro-benzyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a stirred solution of diisopropyl amine (1.3 mL, 9.0 mmol) inTHF (20 mL) was added n-BuLi (1.6 M solution in hexanes, 5.6 mL, 9.0mmol) at 0° C. The reaction was stirred at 0° C. for 15 minutes and thencooled to −78° C. A solution of 3-tert-butoxycarbonylamino-propionicacid tert-butyl ester (1.0 g, 4.1 mmol) in THF (5 mL) was addeddropwise. The mixture was stirred at −78° C. for 2 hours. A solution of4-Bromo-1-bromomethyl-2-fluoro-benzene (1.3 g, 4.9 mmol) in THF (4 mL)was added dropwise. After completion, the dry-ice bath was removed andthe reaction was warmed to 0° C. in an ice bath. After stirring at 0° C.for 30 minutes, the reaction was poured into saturated NH₄Cl aqueoussolution. The organic layer was separated. The aqueous layer wasextracted with EtOAc. The combined organic layers were washed withbrine, dried and concentrated. The residue was purified by columnchromatography (hexane:EtOAc, 20:1 to 5:1) to give2-(4-Bromo-2-fluoro-benzyl)-3-tert-butoxycarbonylamino-propionic acidtert-butyl ester (1.35 g, 77%) as a colorless oil. ¹H NMR (CDCl₃, 400MHz) δ 7.20 (d, J=8.0 Hz, 2H), 7.08 (t, J=8.0 Hz, 1H), 4.86 (m, 1H),3.28 (m, 2H), 2.92 (m, 3H), 1.43 (s, 9H), 1.36 (s, 9H).

Step 2: 2-(4-Bromo-2-fluoro-benzyl)-3-tert-butoxycarbonylamino-propionicacid tert-butyl ester (1.30 g, 3.01 mmol) was dissolved in THF (12 mL)and MeOH (12 mL). A solution of LiOH monohydrate (0.50 g, 12.0 mmol) inH₂O (12 mL) was added. The mixture was heated at reflux overnight. Aftercooling, the solvents were evaporated in vacuo. The residue wasdissolved in water and extracted with ether (2×). The aqueous phase wasacidified with 1N HCl and extracted with EtOAc. The combined organiclayers were washed with brine, dried and concentrated to give2-(4-Bromo-2-fluoro-benzyl)-3-tert-butoxycarbonylamino-propionic acid(1.00 g, 88%) as a white solid. ¹H NMR (CD₃OD, 400 MHz) δ 7.26 (m, 2H),7.19 (m, 1H), 3.27 (m, 2H), 2.86 (m, 3H), 1.43 (s, 9H).

Step 3:3-Amino-2-(4-bromo-2-fluoro-benzyl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by substituting5-piperazin-1-yl-1H-indazole with4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride andsubstituting (D)-Boc-4-chlorophenylalanine with2-(4-Bromo-2-fluoro-benzyl)-3-tert-butoxycarbonylamino-propionic acid inExample 34, Step 2, then removing the Boc protecting group as describedin Example 34, Step 3. ¹H NMR (CD₃OD, 400 MHz) δ 8.37 (s, 1H), 7.40 (d,J=3.6 Hz, 1H), 7.36 (m, 1H), 7.31 (m, 1H), 7.24 (t, J=8.0 Hz, 1H), 6.95(d, J=3.6 Hz, 1H), 4.14 (m, 2H), 4.02 (m, 1H), 3.87 (m, 4H), 3.55 (m,2H), 3.34 (m, 1H), 3.11 (m, 1H), 2.98 (m, 2H). LCMS (APCI+) m/z 461, 463[M+H]⁺; Rt=1.79 min.

Example 79

Preparation of2-(R)-Amino-3-(4-chlorophenyl)-1-(4-imidazo[1,2-a]pyridin-3-yl-piperazin-1-yl)-propan-1-onedihydrochloride

Step 1: A mixture of tert-butyl 1-piperazinecarboxylate (7.45 g, 40.0mmol) and benzotriazole (4.76 g, 40.0 mmol) in H₂O (200 mL) was stirredfor 1 hour. Glyoxal (40 wt. % in water, 2.90 g, 20 mmol) was then addedand the mixture was stirred for 12 hours to produce a white precipitate.The precipitate was filtered off and washed with H₂O. The solids weredissolved in DCM, dried (Na₂SO₄), filtered, and concentrated in vacuo togive 1,2-(benzotriazol-1-yl)-1,2-(4-piperazine-1-carboxylic acidtert-butyl ester)ethane off-white solid (10.0 g).

Step 2: A mixture of 2-aminopyridine (282 mg, 3.00 mmol) and1,2-(benzotriazol-1-yl)-1,2-(4-piperazine-1-carboxylic acid tert-butylester)ethane (1.90 g, 3.00 mmol) in dichloroethane (30 mL) was refluxedfor 3 hours. Powdered KOH (555 mg, 9.90 mmol) was then added, and themixture was stirred for 12 hours. The mixture was filtered and thefiltrate was concentrated in vacuo. The resulting material waschromatographed (SiO₂) using DCM followed by 5% MeOH/DCM as eluent togive 4-imidazo[1,2-a]pyridin-3-yl-piperazine-1-carboxylic acidtert-butyl ester (900 mg). ¹H NMR (CDCl₃, 400 MHz) δ 7.96 (m, 1H), 7.52(m, 1H), 7.25 (m, 1H), 7.10 (m, 1H), 6.78 (m, 1H), 3.61 (m, 4H), 2.98(m, 4H), 1.47 (s, 9H).

Step 3: A solution of4-imidazo[1,2-a]pyridin-3-yl-piperazine-1-carboxylic acid tert-butylester (950 mg, 3.14 mmol) in DCM (10 mL) and 2.0 N HCl in Et₂O (5 mL)was stirred for 12 hours. A precipitate formed and was filtered off togive 3-piperazin-1-yl-imidazo[1,2-a]pyridine dihydrochloride as a redsolid (800 mg).

Step 4: Triethylamine (0.30 mL, 2.18 mmol) was added to a solution of(R)—N-Boc-4-chlorophenylalanine (392 mg, 1.31 mmol),3-piperazin-1-yl-imidazo[1,2-a]pyridine dihydrochloride (300 mg, 1.09mmol), and 1-hydroxybenzotriazole (177 mg, 1.31 mmol) in DMF (5 mL)followed by the addition of1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (251 mg,1.31 mmol) in a single portion. After being stirred for 12 hours, themixture was diluted with H₂O, basified to pH 12 with 1.0 M NaOH, andextracted with DCM. The DCM extracts were dried (Na₂SO₄), filtered, andconcentrated in vacuo. The residue was chromatographed (SiO₂) usingDCM:Et₂O (1:1) followed by 10% MeOH/DCM as eluent to give(R)-[1-(4-chlorobenzyl)-2-(4-imidazo[1,2-a]pyridin-3-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester (350 mg).

Step 5: A solution of(R)-[1-(4-chlorobenzyl)-2-(4-imidazo[1,2-a]pyridin-3-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester (350 mg, 0.72 mmol) in DCM (3 mL) and 2.0 N HCl inEt₂O (2 mL) was stirred for 12 hours. The mixture was concentrated invacuo and the resulting material was chromatographed (SiO₂) using 10%MeOH/DCM followed by 10% (7 N NH₃ in MeOH)/DCM as eluent. This materialwas dissolved in MeOH followed by the addition of 2.0 N HCl in Et₂O, andthen concentrated in vacuo. The resulting gum was stripped fromisopropyl alcohol, and then from acetonitrile to give2-(R)-amino-3-(4-chlorophenyl)-1-(4-imidazo[1,2-a]pyridin-3-yl-piperazin-1-yl)-propan-1-onedihydrochloride as a white solid (100 mg). ¹H NMR (DMSO-d₆, 400 MHz) δ8.66 (d, J=6.8 Hz, 1H), 8.49 (bs, 3H), 7.89 (m, 3H), 7.45 (m, 1H), 7.38(d, J=8.3 Hz, 2H), 7.26 (d, J=8.3 Hz, 2H), 4.69 (bs, 1H), 3.49 (m, 6H),3.08 (m, 1H), 2.94 (m, 3H), 2.85 (m, 1H). LCMS (APCI+) m/z 384, 386[M+H]⁺.

Example 80

Preparation of2(R)-Amino-3-(4-chlorophenyl)-3-methyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-one

Step 1: The 2-(4-chlorophenyl)-2-methyl-propionic acid (6.10 g, 30.7mmol) was dissolved in 120 mL of dry THF at room temperature. A 70% w/wsolution of Red-Al (28.25 mL, 0.101 mol) was added dropwise via syringeover 5 minutes (vigrous bubbling). The mixture was heated to reflux forthree hours. The solution was cooled to 0 C and carefully quenched withthe addition of saturated sodium tartrate solution (100 mL, violenthydrogen evolution) and 100 mL of water. The aqueous mixture wasextracted with ethyl acetate, and the combined organic was washed withdiluted NaHCO₃ solution, then brine, separated, dried over MgSO₄,filtered, and concentrated in vacuo to afford2-(4-chlorophenyl)-2-methyl-propan-1-ol as a colorless oil (5.70 g,99%). The material was used without purification. ¹H NMR (CDCl₃, 400MHz) δ 7.32 (s, 5H), 3.62 (d, J=4.4 Hz, 2H), 1.33 (s, 6H), 1.23 (t,J=4.4 Hz, 1H).

Step 2: The DMSO (436 mL, 61.4 mmol) was dissolved in 100 mL of DCM andtreated with oxalyl chloride (4.02 μL, 46.6 mmol) at −78° C. Thesolution stirred for 30 minutes at −78° C. before the2-(4-chlorophenyl)-2-methyl-propan-1-ol (5.67 g, 30.7 mmol) was addeddropwise as a solution in 10 mL of DCM. After addition was complete, thesolution was stirred for two hours at −78° C., and then treated withtriethyl amine (25.7 mL, 184 mmol). The solution was allowed to warm toambient temperature and stir for three hours. The solution was quenchedwith the addition of water and partitioned with more DCM. The aqueouswas extracted with DCM, and the combined organics were dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bychromatography (silica, hexanes/ethyl acetate gradients) to afford2-(4-chlorophenyl)-2-methylpropionaldehyde as a yellow oil (5.60 g,99%). ¹H NMR (CDCl₃, 400 MHz) δ 9.48 (s, 1H), 7.35 (d, J=8.8 Hz, 2H),7.21 (d, J=8.8 Hz, 1H), 1.45 (s, 6H).

Step 3: The 2-(4-chlorophenyl)-2-methylpropionaldehyde (5.60 g, 30.7mmol) and (S)-4-methyl-benzenesulfinic acid amide (5.00 g, 32.2 mmol)were dissolved in 300 mL of DCM and treated with Ti(OEt)₄ (32.1 mL, 153mmol). The mixture was heated to reflux under nitrogen for four hours.The solution was cooled in an ice bath and quenched with thedropwise-addition of 200 mL of water. The resulting precipitate (Tisalts) were removed by filtration through a plug of celite and washedwith DCM. The resulting filtrate was separated, and the aqueous wasextracted with more DCM. The combined organic was dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified byfiltration through a plug of silica gel (hexanes:ethyl acetate, 1:1) toafford the (R)-4-methylbenzenesulfinic acid[2-(4-chlorophenyl)-2-methylpropylidene]-amide as a colorless oil, whichsolidified upon standing to give a white solid (9.28 g, 95%). ¹H NMR(CDCl₃, 400 MHz) δ 8.14 (s, 1H), 7.54 (d, J=8.4 Hz, 2H), 7.31 (d, J=8.8Hz, 2H), 7.27 (d, J=8.8 Hz, 2H), 7.14 (m, 2H), 2.42 (s, 3H), 1.52 (s,3H), 1.47 (s, 3H).

Step 4: The diethyl aluminum cyanide (43.5 mL of a 1.0M solution intoluene, 43.5 mmol) was added to isopropanol (28.9 mL, 377 mmol) andstirred at 10° C. for 15 minutes. This solution was cannulated into the(R)-4-methylbenzenesulfinic acid[2-(4-chlorophenyl)-2-methyl-propylidene]-amide (9.28 g, 29.0 mmol) as asolution in 290 mL of THF at −78° C. This solution was allowed to stirfor 15 minutes at −78° C. then allowed to warm slowly to roomtemperature overnight. The solution was quenched with the addition ofdiluted NaHCO₃ solution and extracted with ethyl acetate. The combinedorganic was washed with brine, separated, dried over Na₂SO₄, filtered,and concentrated in vacuo to afford the 4-methyl-benzenesulfinic acid[2-(4-chlorophenyl)-1-cyano-2-methyl-propyl]-amide as a colorless oil(9.55 g, 95% yield). The material was heated to 110° C. (reflux) inconcentrated HCl solution over the weekend. The solution was cooled toroom temperature, washed with ether, then concentrated in vacuo to givethe (S)-2-amino-3-(4-chlorophenyl)-3-methyl-butyric acid hydrochloridesalt a white solid (1.61 g, 21%). ¹H NMR (DMSO-d₆, 400 MHz) δ 8.19 (brs,1H), 7.48 (d, J=8.0 Hz, 2H), 7.41 (d, J=8.0 Hz, 2H), 7.13 (s, 3H), 4.15(s, 1H), 1.42 (s, 3H), 1.40 (s, 3H). LCMS (APCI+) m/z 228 [M+H]⁺;Rt=1.81 min.

Step 5: The (S)-2-amino-3-(4-chlorophenyl)-3-methyl-butyric acidhydrochloride salt (1.00 g, 3.79 mmol) was dissolved in 6 mL of 2M NaOHsolution at room temperature and treated with di-tert-butyl di-carbonate(957 L, 4.16 mmol). The solution was allowed to stir for four hours tocompletion, and the aqueous solution was acidified with the addition of1M HCl solution (pH=2-3). The aqueous was extracted with ethyl acetate,and the organics were combined. The organic was washed with brine,separated, dried over MgSO₄, filtered, and concentrated in vacuo to givea colorless oil. The residue was purified by chromatography (silica,hexanes/ethyl acetate gradients) to afford the(S)-2-tert-butoxycarbonylamino-3-(4-chlorophenyl)-3-methyl-butyric acid(636 mg, 51%). ¹H NMR (CDCl₃, 400 MHz) δ 12.67-10.55 (brs, 1H), 7.29 (m,4H), 4.97 (d, J=8.4 Hz, 1H), 4.55 (d, J=8.4 Hz, 1H), 1.45 (s, 9H), 1.38(s, 6H). LCMS (APCI+) m/z 228 [M-Boc+H]⁺; Rt=3.20 min.

Step 6: The 4-piperazin-1-yl-quinazoline bis-hydrochloride (220 mg,0.766 mmol),(S)-2-tert-butoxycarbonylamino-3-(4-chlorophenyl)-3-methyl-butyric acid(251 mg, 0.766 mmol, 1.0 equiv), 1-hydroxybenzotriazole (109 mg, 0.804mmol, 1.05 equiv), and EDCI (154 mg, 0.804 mmol, 1.05 equiv) weredissolved/suspended in 6.0 mL of DMF. The mixture was treated withtriethylamine (427 μL, 3.06 mmol) and allowed to stir overnight tocompletion. The reaction was partitioned between ethyl acetate anddiluted NaHCO₃ solution. The aqueous was extracted with ethyl acetate,and the organics were combined. The organic was washed with water, thenbrine, separated, dried over MgSO₄, filtered, and concentrated in vacuo.The residue was eluted through a small plug of silica gel with ethylacetate and concentrated in vacuo. The protected intermediate wasimmediately dissolved in 1 mL of dioxane and treated with 4M HCl indioxane (1.92 mL, 7.66 mmol) at room temperature for four hours. Theresulting precipitate was triturated with ether, then filtered to affordthe(R)-2-amino-3-(4-chlorophenyl)-3-methyl-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-onebis-hydrochloride as a light-yellow solid (277 mg, 73%) upon dryingunder vacuum. ¹H NMR (DMSO-d₆, 400 MHz) δ 8.89 (s, 1H), 8.46 (brs, 3H),8.17 (d, J=8.4 Hz, 1H), 8.06 (t, J=7.6 Hz, 1H), 7.97 (d, J=8.4 Hz, 1H),7.74 (d, J=8.0 Hz, 1H), 7.50 (d, J=8.0 Hz, 2H), 7.42 (d, J=8.0 Hz, 2H),4.53 (brs, 1H), 4.09 (m, 3H), 3.90 (m, 3H), 3.32 (m, 1H), 3.04 (t, J=9.2Hz, 1H), 1.51 (s, 3H), 1.43 (s, 3H). LCMS (APCI+) m/z 424 [M+H]⁺;Rt=1.90 min.

Example 81

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluoren-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1:4-(6,7,8,9-Tetrahydro-5H-1,3,9-triaza-fluoren-4-yl)-piperazine-1-carboxylicacid tert-butyl ester was prepared by the procedures described inExample 40, Step 1, substituting 4-chloro-5-iodopyrimidine with4-Chloro-6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluorene (prepared from2-Amino-1-(4-methoxy-benzyl)-4,5,6,7-tetrahydro-1H-indole-3-carbonitrileaccording to the literature: Traxler, P. M. et. al. (1996), J. Med.Chem., 39, 2285-2292). ¹H NMR (CDCl₃, 400 MHz) δ 10.24 (s, 1H), 8.32 (s,1H), 3.59 (m, 4H), 3.55 (m, 4H), 2.78 (m, 4H), 1.92 (m, 2H), 1.82 (m,2H), 1.43 (s, 9H). LCMS (APCI+) m/z 358 [M+H]⁺; Rt=3.12 min.

Step 2: 4-piperazin-1-yl-6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluorenedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-Chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with4-(6,7,8,9-Tetrahydro-5H-1,3,9-triaza-fluoren-4-yl)-piperazine-1-carboxylicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.42 (s, 1H), 4.14 (m,4H), 3.48 (m, 4H), 2.83 (m, 4H), 1.96 (m, 2H), 1.86 (m, 2H). LCMS(APCI+) m/z 258 [M+H]⁺; Rt=1.52 min.

Step 3: To a suspension of4-piperazin-1-yl-6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluorenedihydrochloride (20 mg, 0.061 mmol) and (D)-Boc-4-chlorophenylalanine(20 mg, 0.067 mmol) were added DIEA (63 μL, 0.36 mmol) and HBTU (25 mg,0.067 mmol). The reaction was stirred at room temperature for 2 hours.The mixture was partitioned between water and EtOAc. The organic layerwas washed with aqueous NaHCO₃ and brine, dried and concentrated. Theresidue was purified by column chromatography (DCM:MeOH, 40:1 to 20:1)to give(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluoren-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester as a colorless oil. Removal of the Boc protectinggroup by procedures described in Example 34, Step 3 afforded(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluoren-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (28 mg, 90%) as a white solid. ¹H NMR (CD₃OD, 400 MHz) δ8.29 (s, 1H), 7.42 (d, J=8.0 Hz, 2H), 7.33 (d, J=8.0 Hz, 2H), 4.74 (m,1H), 3.75 (m, 6H), 3.38 (m, 1H), 3.16 (m, 3H), 2.77 (m, 4H), 1.92 (m,4H) LCMS (APCI+) m/z 439, 441 [M+H]⁺; Rt=2.16 min.

Example 82

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(quinazolin-4-yloxy)-piperidin-1-yl]-propan-1-one

Step 1: To a stirred suspension of NaH (60%, 0.146 g, 3.65 mmol) in DMF(15 mL) was added dropwise a solution of4-Hydroxypiperidine-1-carboxylic acid tert-butyl ester (0.611 g, 3.04mmol) in DMF (5 mL) at 0° C. The reaction was stirred for 1 hour andthen 4-chloroquinazoline (0.500 g, 3.04 mmol) was added. The mixture wasallowed to warm to room temperature and stirred overnight. The mixturewas partitioned between H₂O and EtOAc. The aqueous phase was extractedwith EtOAc. The combined organic layers were washed with brine, driedand concentrated. The residue was purified by column chromatography(hexanes:EtOAc, 2:1) to give4-(Quinazolin-4-yloxy)-piperidine-1-carboxylic acid tert-butyl ester(0.76 g, 76%) as a colorless oil. Removal of the Boc group by theprocedures described in Example 34, Step 3 afforded4-(Piperidin-4-yloxy)-quinazoline dihydrochloride as a white solid. ¹HNMR (CD₃OD, 400 MHz) δ 9.35 (s, 1H), 8.59 (d, J=8.4 Hz, 1H), 8.31 (td,J=7.2 Hz, J=1.2 Hz 1H), 8.11 (d, J=8.4 Hz, 1H), 8.04 (t, J=7.2 Hz, 1H),6.00 (m, 1H), 3.55 (m, 2H), 3.40 (m, 2H), 2.42 (m, 4H). LCMS (APCI+) m/z230 [M+H]⁺; Rt=1.67 min.

Step 2:(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(quinazolin-4-yloxy)-piperidin-1-yl]-ethyl}-carbamicacid 9H-fluoren-9-ylmethyl ester was prepared by the proceduresdescribed in Example 34, Step 2, substituting5-piperazin-1-yl-1H-indazole with 4-(Piperidin-4-yloxy)-quinazolinedihydrochloride and substituting (D)-Boc-4-chlorophenylalanine with(D)-Fmoc-4-chlorophenylalanine. ¹H NMR (CDCl₃, 400 MHz) δ 8.77 (s, 1H),8.12 (m, 1H), 8.02 (s, 1H), 7.94 (m, 1H), 7.84 (m, 1H), 7.77 (m, 2H),7.59 (m, 4H), 7.41 (m, 2H), 7.30 (m, 4H), 7.15 (m, 2H), 5.71 (d, J=8.4Hz, 1H), 5.55 (m, 1H), 4.94 (m, 1H), 4.40 (m, 2H), 4.21 (m, 1H), 3.85(m, 1H), 3.61 (m, 1H), 3.38 (m, 1H), 3.02 (m, 2H), 1.94 (m, 5H). LCMS(APCI+) m/z 633, 635 [M+H]⁺; Rt=3.98 min.

Step 3: To a stirred solution of(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(quinazolin-4-yloxy)-piperidin-1-yl]-ethyl}-carbamicacid 9H-fluoren-9-ylmethyl ester (0.166 g, 0.262 mmol) in DCM (5 mL) wasadded piperidine (1 mL). The reaction was stirred at room temperaturefor 4 hours. The volatiles were evaporated. The residue was purified bycolumn chromatography (DCM:MeOH, 50:1 to 10:1) to give(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(quinazolin-4-yloxy)-piperidin-1-yl]-propan-1-one(0.099 g, 92%) as a colorless oil. ¹H NMR (CDCl₃, 400 MHz) δ 8.77 (s,1H), 8.15 (m, 1H), 7.94 (d, J=8.4 Hz, 1H), 7.85 (m, 1H), 7.59 (m, 1H),7.30 (d, J=8.0 Hz, 2H), 7.17 (d, J=8.0 Hz, 2H), 5.57 (m, 1H), 3.95 (m,2H), 3.10-3.70 (m, 3H), 2.95 (m, 1H), 2.82 (m, 1H), 1.30-2.10 (m, 4H).LCMS (APCI+) m/z 411, 413 [M+H]⁺; Rt=2.19 min.

Example 83

Preparation of4-Amino-2-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-onedihydrochloride

Step 1: An analogous reaction to that described in example 61 steps 1-3,but starting with (3,4-dichlorophenyl)-acetic acid methyl ester yielded4-tert-butoxycarbonylamino-2-(3,4-dichlorophenyl)-butyric acid. ¹H NMR(CDCl₃, 400 MHz) δ 7.41 (m, 2H), 7.17 (d, J=8 Hz, 1H), 4.61 (brs, 1H),3.58 (m, 1H), 3.16 (m, 2H), 2.28 (m, 1H), 1.92 (m, 1H), 1.44 (s, 9H).

Step 2: The 4-piperazin-1-yl-quinazoline (20 mg, 0.070 mmol) wasdissolved in 1 mL CHCl₃ and4-tert-butoxycarbonylamino-2-(3,4-dichlorophenyl)-butyric acid (36 mg,0.10 mmol) was added. PS-carbodiimide resin (0.21 mmol) was added andthe mixture was shaken overnight. The reaction mixture was filtered andthe filtrate was concentrated in vacuo. The residue was purified bycolumn chromatography (silica gel eluted with 1:4 DCM/EtOAc) to affordthe pure Boc-protected intermediate. The material was dissolved in 1.0mL of 1,4-dioxane and treated with 1.0 mL of 4M HCl in dioxane (4 mmol).The solution was allowed to stir at room temperature overnight tocompletion. The reaction mixture was concentrated to dryness and theresidue was triturated with 2 mL diethyl ether. The solid was filteredunder a nitrogen atmosphere and was allowed to dry under vacuum for 2hours to afford4-amino-2-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-onedihydrochloride (15 mg, 42%). ¹H NMR (CD₃OD, 400 MHz) δ 8.69 (s, 1H),8.22 (d, J=9 Hz, 1H), 8.04 (t, J=8 Hz, 1H), 7.77 (m, 2H), 7.54 (m, 2H),7.31 (d, J=8.6 MHz, 1H), 4.30 (m, 4H), 3.98 (m, 4H), 3.60 (m, 1H), 2.98(m, 1H), 2.86 (m, 1H), 2.36 (m, 1H), 2.02 (m, 1H).

Example 84

Preparation of2-(3,4-Dichlorophenyl)-4-methylamino-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-onedihydrochloride

2-(3,4-dichlorophenyl)-4-Boc-amino-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-onewas N-methylated following a literature procedure (Mahavir Prashad etal. Org. Lett. 2003, 5(2), 125-128) to give[3-(3,4-Dichlorophenyl)-4-oxo-4-(4-quinazolin-4-yl-piperazin-1-yl)-butyl]-methyl-carbamicacid tert-butyl ester, which was treated with excess HCl/dioxane tofurnish2-(3,4-dichlorophenyl)-4-methylamino-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-onedihydrochloride (7 mg, 31%). ¹H NMR (DMSO-d₆, 400 MHz) δ 8.85 (br s,2H), 8.84 (s, 1H), 8.17 (d, J=7.6 Hz, 1H), 8.01 (m, 1H), 7.89 (d, J=7.6Hz, 1H), 7.69-7.61 (m, 3H), 7.33 (d, J=7.6 Hz, 1H), 4.40 (m, 1H), 4.11(br s, 3H), 3.95 (m, 2H), 3.78 (m, 1H), 3.73 (m, 2H), 3.45 (m, 3H), 2.80(m, 1H), 2.71 (m, 1H), 2.25 (m, 1H), 1.97 (m, 1H). LCMS (APCI+) m/z 458[M+H]⁺. HPLC R_(t)=1.78 min.

Example 85

Preparation of1-{4-[4-Amino-2-(3,4-dichlorophenyl)-butyryl]-piperazin-1-yl}-quinolizin-4-onehydrochloride

Step 1: To a solution of 4-Pyridin-2-ylmethyl-piperazine-1-carboxylicacid tert-butyl ester (prepared from 1-Pyridin-2-ylmethyl-piperazineaccording to the literature: J. Med. Chem. (1993), 36, 2984) (2.00 g,7.21 mmol) in THF (15 mL) was added n-BuLi (1.6 M in hexanes, 5.0 mL,7.9 mmol) at −78° C. The mixture was allowed to warm to room temperatureand stirred for 30 minutes. The solution was then cooled to −78° C. anda solution of diethyl ethoxymethylenemalonate (1.72 g, 7.93 mmol) in THF(2 mL) was added dropwise. The reaction mixture was allowed to warm to0° C. over 1 hour and stirred at 0° C. for 1 hour. The reaction waspoured into water and extracted with EtOAc. The combined organic layerswere washed with brine, dried and concentrated. The residue was purifiedby column chromatography (hexanes:EtOAc, 8:1 to 1;1) to give2-[2-(4-tert-Butoxycarbonyl-piperazin-1-yl)-1-ethoxy-2-pyridin-2-yl-ethyl]-malonicacid diethyl ester (2.40 g, 67%) as a mixture of diastereomers.Diastereomer 1: ¹H NMR (CDCl₃, 400 MHz) δ 8.55 (d, J=4.4 Hz, 1H), 7.63(td, J=7.6 Hz, J=1.6 Hz, 1H), 7.21 (d, J=7.6 Hz, 1H), 7.16 (dd, J=7.6Hz, J=1.6 Hz, 1H), 4.74 (m, 1H), 4.16 (m, 5H), 3.75 (q, J=7.2 Hz, 2H),3.52 (d, J=4.4 Hz, 1H), 3.38 (m, 4H), 2.67 (m, 2H), 2.44 (m, 2H), 1.40(s, 9H), 1.22 (m, 6H), 1.44 (t, J=7.2 Hz, 3H). LCMS (APCI+) m/z 494[M+H]⁺; Rt=3.61 min. Diastereomer 2: ¹H NMR (CDCl₃, 400 MHz) δ 8.58 (d,J=4.8 Hz, 1H), 7.59 (td, J=7.6 Hz, J=1.6 Hz, 1H), 7.15 (dd, J=7.6 Hz,J=1.6 Hz, 1H), 7.08 (d, J=8.0 Hz, 1H), 4.91 (m, 1H), 4.16 (m, 5H), 3.53(m, 1H), 3.22 (m, 6H), 2.50 (m, 2H), 2.19 (m, 2H), 1.34 (s, 9H), 1.24(m, 6H), 0.69 (t, J=7.2 Hz, 3H). LCMS (APCI+) m/z 494 [M+H]⁺; Rt=3.62min.

Step 2:2-[2-(4-tert-Butoxycarbonyl-piperazin-1-yl)-1-ethoxy-2-pyridin-2-yl-ethyl]-malonicacid diethyl ester (2.40 g, 4.86 mmol) was dissolved in xylene (20 mL)and heated at 140° C. for 12 hours. After cooling, the volatiles wereevaporated and the residue was purified by column chromatography (EtOAc)to give1-(4-tert-Butoxycarbonyl-piperazin-1-yl)-4-oxo-1,9a-dihydro-4H-quinolizine-3-carboxylicacid ethyl ester (1.39 g, 71%) as an orange solid. ¹H NMR (CDCl₃, 400MHz) δ 9.48 (d, J=7.2 Hz, 1H), 8.34 (s, 1H), 7.69 (td, J=7.2 Hz, J=1.2Hz, 1H), 7.23 (m, 1H), 4.44 (q, J=7.2 Hz, 2H), 4.14 (m, 2H), 3.10 (m,2H), 2.88 (m, 4H), 1.51 (s, 9H), 1.43 (t, J=7.2 Hz, 3H). LCMS (APCI+)m/z 402 [M+H]⁺; Rt=2.85 min.

Step 3: A mixture of1-(4-tert-Butoxycarbonyl-piperazin-1-yl)-4-oxo-1,9a-dihydro-4H-quinolizine-3-carboxylicacid ethyl ester (0.320 g, 0.797 mmol) in concentrated HCl (5 mL) wasrefluxed for 30 minutes. After cooling, the reaction was basified withaqueous NaHCO₃ solution and thoroughly extracted with DCM. The combinedorganic layers were washed with brine, dried and concentrated to give1-piperazin-1-yl-1,9a-dihydro-quinolizin-4-one (0.075 g, 41%) as ayellow oil. ¹H NMR (CDCl₃, 400 MHz) δ 9.16 (d, J=7.2 Hz, 1H), 8.20 (d,J=9.2 Hz, 1H), 7.71 (d, J=9.6 Hz, 1H), 7.37 (t, J=7.2 Hz, 1H), 7.03 (t,J=6.4 Hz, 1H), 6.61 (d, J=9.2 Hz, 1H), 3.07 (m, 4H), 2.87 (m, 4H). LCMS(APCI+) m/z 230 [M+H]⁺; Rt=0.29 min.

Step 4:1-{4-[4-Amino-2-(3,4-dichlorophenyl)-butyryl]-piperazin-1-yl}-quinolizin-4-onehydrochloride was prepared by substituting 5-piperazin-1-yl-1H-indazolewith 1-piperazin-1-yl-1,9a-dihydro-quinolizin-4-one and substituting(D)-Boc-4-chlorophenylalanine with4-tert-Butoxycarbonylamino-2-(3,4-dichlorophenyl)-butyric acid inExample 34, Step 2, then removing the Boc protecting group as describedin Example 34, Step 3. ¹H NMR (CD₃OD, 400 MHz) δ 9.25 (d, J=7.2 Hz, 1H),9.57 (d, J=8.8 Hz, 1H), 8.00 (m, 2H), 7.71 (d, J=7.2 Hz, 1H), 7.58 (m,2H), 7.33 (d, J=8.0 Hz, 1H), 7.02 (d, J=8.4 Hz, 1H), 4.31 (m, 1H), 3.82(m, 5H), 3.65 (m, 2H), 3.34 (m, 1H), 2.98 (m, 1H), 2.86 (m, 1H), 2.36(m, 1H), 2.03 (m, 1H). LCMS (APCI+) m/z 459, 461, 463 [M+H]⁺; Rt=2.02min.

Example 86

Preparation of(2R)-2-Amino-3-phenyl-1-(4-quinazolin-4-yl-piperidin-1-yl)-propan-1-onedihydrochloride

Step 1: To a 25 mL flask was charged4-Trifluoromethanesulfonyloxy-3,6-dihydro-2H-pyridine-1-carboxylic acidbenzyl ester (prepared from 4-Oxo-piperidine-1-carboxylic acid benzylester according to the literature: Wustrow, D. J. et. al. (1991),Synthesis, 993-995. 1.14 g, 3.12 mmol), 4-chloroquinazoline (0.512 g,3.12 mmol), lithium chloride (0.397 g, 9.36 mmol), Pd(PPh₃)₄ (0.180 g,0.156 mmol) and hexamethyl ditin (1.02 g, 3.12 mmol). 1,4-Dioxane (20mL) was added and the reaction was degassed with N₂ for 15 minutes. Themixture was stirred at reflux overnight. After cooling, the blacksuspension was poured into saturated aqueous potassium fluoridesolution. The mixture was diluted with EtOAc and stirred for 2 hours.The organic phase was separated. The aqueous phase was extracted withEtOAc. The combined organic layers were washed with brine, dried andconcentrated. The residue was purified by column chromatography(hexanes:EtOAc, 2:3) to give4-Quinazolin-4-yl-3,6-dihydro-2H-pyridine-1-carboxylic acid benzyl ester(0.790 g, 73%) as a colorless oil. ¹H NMR (CDCl₃, 400 MHz) δ 9.27 (s,1H), 8.19 (d, J=7.2 Hz, 1H), 8.07 (d, J=8.8 Hz, 1H), 7.91 (m, 1H), 7.62(t, J=7.2 Hz, 1H), 7.41 (m, 6H), 6.18 (m, 1H), 5.23 (s, 2H), 4.32 (s,2H), 3.85 (t, J=5.6 Hz, 1H), 2.80 (br s, 2H). LCMS (APCI+) m/z 346[M+H]⁺; Rt=3.16 min.

Step 2: To a stirred solution of4-Quinazolin-4-yl-3,6-dihydro-2H-pyridine-1-carboxylic acid benzyl ester(0.907 g, 2.63 mmol) in MeOH (30 mL) under N₂ was cautiously added 10%Pd on carbon (100 mg). The reaction was hydrogenated at 50 psi using aparr shaker for 3 days. The catalyst was removed by filtration. Thefiltrate was evaporated under vacuum. The resulting residue was purifiedby column chromatography (DCM:MeOH, 20:1 to DCM:MeOH:Et₃N, 100:10:1) togive 4-(1,2,3,6-Tetrahydro-pyridin-4-yl)-quinazoline (0.358 g, 64%) as awhite waxy solid. ¹H NMR (CDCl₃, 400 MHz) δ 9.27 (s, 1H), 8.19 (d, J=8.4Hz, 1H), 8.06 (d, J=8.4 Hz, 1H), 7.88 (t, J=8.4 Hz, 1H), 7.65 (t, J=8.4Hz, 1H), 3.71 (m, 1H), 3.31 (m, 2H), 2.90 (m, 2H), 1.99 (m, 4H). LCMS(APCI+) m/z 214 [M+H]⁺; Rt=1.60 min.

Step 3:(2R)-[1-Benzyl-2-oxo-2-(4-quinazolin-4-yl-piperidin-1-yl)-ethyl]-carbamicacid tert-butyl ester was prepared by substituting5-piperazin-1-yl-1H-indazole with4-(1,2,3,6-Tetrahydro-pyridin-4-yl)-quinazoline and substituting(D)-Boc-4-chlorophenylalanine with (D)-Boc-phenylalanine in Example 34,Step 2. ¹H NMR (CDCl₃, 400 MHz) (1:1 mixture of rotamers) δ 9.26, 9.21(2s, 1H, rotamers), 8.08 (m, 2H), 7.89 (m, 1H), 7.65 (m, 1H), 7.28 (m,5H), 5.51, 5.43 (2d, J=8.4 Hz, 1H, rotamers), 4.92 (m, 1H), 4.72 (m,1H), 3.95, 3.83 (2d, J=13.2 Hz, 1H, rotamers), 3.65 (m, 1H), 2.20-3.30(m, 4H), 1.20-2.10 (m, 4H), 1.44, 1.42 (2s, 9H, rotamers). LCMS (APCI+)m/z 461 [M+H]⁺; Rt=3.23 min.

Step 4:(2R)-2-Amino-3-phenyl-1-(4-quinazolin-4-yl-piperidin-1-yl)-propan-1-onedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-Chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with(2R)-[1-Benzyl-2-oxo-2-(4-quinazolin-4-yl-piperidin-1-yl)-ethyl]-carbamicacid tert-butyl ester. LCMS (APCI+) m/z 361 [M+H]⁺; Rt=2.38 min.

Example 87

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,6-dihydro-2H-pyridin-1-yl]-propan-1-onedihydrochloride

Step 1: To a nitrogen flushed flask containing4-(4,4,5,5-Tetramethyl-[1,3,2]dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester (prepared fromtert-butyl-4-oxopiperidine-1-carboxylate according to the literature:Eastwood, P. R. (2000), Tetrahedron Lett., 3705-3708. 127 mg, 0.410mmol), K₂CO₃ (142 mg, 1.02 mmol) anddichloro[1,1′-bis(diphenylphosphino)-ferrocene]palladium(II)dichloromethane adduct (17 mg, 0.020 mmol) was added a solution of1-Benzenesulfonyl-4-chloro-1H-pyrrolo[2,3-b]pyridine (100 mg, 0.342mmol) in DMF (3 mL). The mixture was heated at 80° C. for 36 hours. Themixture was cooled to room temperature and partitioned between EtOAc andwater. The combined organic layers were washed with saturated aqueousNaHCO₃ and brine, dried and concentrated. The residue was purified byflash chromatography on silica gel, eluting with hexanes:EtOAc (2:1) togive4-(1-Benzenesulfonyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester (0.068 g, 45%) as a colorless oil. ¹H NMR (CDCl₃,400 MHz) δ 8.37 (d, J=5.2 Hz, 1H), 8.20 (d, J=8.0 Hz, 2H), 7.73 (d,J=4.0 Hz, 1H), 7.56 (m, 1H), 7.49 (m, 2H), 7.03 (d, J=5.2 Hz, 1H), 6.73(d, J=4.0 Hz, 1H), 6.14 (br s, 1H), 4.12 (m, 2H), 3.65 (m, 2H), 2.55 (m,2H), 1.50 (s, 9H). LCMS (APCI+) m/z 440 [M+H]⁺; Rt=3.84 min.

Step 2:1-Benzenesulfonyl-4-(1,2,3,6-tetrahydro-pyridin-4-yl)-1H-pyrrolo[2,3-b]pyridinedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-Chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with4-(1-Benzenesulfonyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-3,6-dihydro-2H-pyridine-1-carboxylicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.36 (d, J=5.2 Hz, 1H),8.15 (d, J=8.0 Hz, 2H), 7.93 (d, J=4.0 Hz, 1H), 7.76 (m, 1H), 7.56 (m,2H), 7.29 (d, J=5.2 Hz, 1H), 6.97 (d, J=4.0 Hz, 1H), 6.29 (br s, 1H),3.93 (m, 2H), 3.60 (m, 2H), 2.85 (m, 2H). LCMS (APCI+) m/z 340 [M+H]⁺;Rt=1.87 min.

Step 3: To a solution of1-Benzenesulfonyl-4-(1,2,3,6-tetrahydro-pyridin-4-yl)-1H-pyrrolo[2,3-b]pyridinedihydrochloride (25 mg, 0.061 mmol) and (D)-Boc-4-chlorophenylalanine(20 mg, 0.067 mmol) in DMF (2 mL) were added DIEA (63 μL, 0.36 mmol) andHBTU (25 mg, 0.067 mmol). The reaction was stirred at room temperaturefor 2 hours. The mixture was partitioned between water and EtOAc. Theorganic layer was washed with aqueous NaHCO₃ and brine, dried andconcentrated. The residue was dissolved in THF (0.3 mL) and MeOH (0.3mL). A solution of lithium hydroxide monohydrate (10 mg, 0.24 mmol) inH₂O (0.3 mL) was added. The mixture was heated at 50° C. overnight.After cooling, the reaction was partitioned between EtOAc and water. Theorganic layer was washed with brine, dried and concentrated. The residuewas purified by column chromatography (hexanes:EtOAc, 1:1 to 3:1) togive(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,6-dihydro-2H-pyridin-1-yl]-ethyl}-carbamicacid tert-butyl ester as a colorless oil. Removal of the Boc group bythe procedures described in Example 34, Step 3 afforded(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(1H-pyrrolo[2,3-b]pyridin-4-yl)-3,6-dihydro-2H-pyridin-1-yl]-propan-1-onedihydrochloride (9 mg, 33%) as a white solid. ¹H NMR (CD₃OD, 400 MHz)(1:1 mixture of rotamers) δ 8.37 (m, 1H), 7.73 (s, 1H), 7.47, 7.40 (2d,J=6.0 Hz, rotamers), 7.33 (m, 3H), 7.23 (d, J=8.0 Hz, 2H), 6.99 (m, 1H),6.33, 6.31 (2s, 1H, rotamers), 4.74 (m, 1H), 4.36 (m, 1H), 4.21 (m, 1H),3.45-3.90 (m, 2H), 3.05-3.30 (m, 2H), 1.90-2.70 (m, 2H). LCMS (APCI+)m/z 381, 383 [M+H]⁺; Rt=1.95 min.

Example 88

Preparation of2(R)-Amino-3-(4-chlorophenyl)-N-(1-quinazolin-4-yl-azetidin-3-yl)-propionamide

Step 1: The (1-benzhydrylazetidin-3-yl)-carbamic acid tert-butyl ester(500 mg, 1.48 mmol), Pd/C (10% w/w, 157 mg, 0.07 mmol), and ammoniumformate (932 mg, 14.8 mmol) were weighed into a 25 RBF equipped with acondenser, degassed 3 times, and suspended/dissolved in 6 mL ofmethanol. The mixture was heated to 60 C for 4 hours to completion andwas allowed to cool to room temperature. The mixture was filteredthrough a plug of celite washed with ethanol, and the filtrate wasconcentrated in vacuo. The residue was re-dissolved in 30 mL of DCM,dried over Na₂SO₄, filtered, and concentrated in vacuo to afford thecrude intermediate. The intermediate and 4-chloroquinazoline (268 mg,1.63 mmol) were dissolved in 6 mL of NMP, then treated withdiisopropylethyl amine (515 μL, 2.96 mmol). The solution was heated to80° C. overnight to completion affording an orange mixture. Aftercooling to room temperature, the solution was diluted with ethyl acetateand poured into diluted NaHCO₃ solution. The aqueous was extracted withethyl acetate, and the organics were combined. The organic was washedwith water, brine, separated, dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was purified by chromatography(silica gel eluted with 9:1 MeOH:EtOAc) to afford the pure(1-quinazolin-4-yl-azetidin-3-yl)-carbamic acid tert-butyl ester as atan solid (390 mg, 88%). ¹H NMR (DMSO-d₆, 400 MHz) δ 8.47 (s, 1H), 7.94(d, J=8.4 Hz, 1H), 7.78 (t, J=7.6 Hz, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.67(m, 1H), 7.48 (t, J=7.6 Hz, 1H), 4.73 (m, 2H), 4.50 (m, 1H), 4.30 (m,2H), 1.41 (s, 9H). LCMS (APCI+) m/z 301 [M+H]⁺; Rt=2.26 min.

Step 2: The (1-quinazolin-4-yl-azetidin-3-yl)-carbamic acid tert-butylester (390 mg, 1.30 mmol) was dissolved in 7 mL of 4M HCl and allowed tostir at 80° C. to completion after three hours. The aqueous solution waswashed with ether (discarded), and the aqueous layer was concentrated invacuo to afford the de-protected intermediate as a white solid. Theflask containing this solid was charged with HOBt (193 mg, 1.43 mmol),EDCI (274 mg, 1.43 mmol), and the2(S)-tert-butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid (389 mg.1.30 mmol). The mixture was suspended/dissolved in 12.0 mL of DMF andtreated with TEA (905 L, 6.49 mmol). The mixture was allowed to stir forfour hours to completion. The contents were partitioned between ethylacetate and diluted NaHCO₃ solution. The aqueous was extracted withethyl acetate, and the organics were combined. The organic was washedwith water, brine, separated, dried over MgSO₄, filtered, andconcentrated in vacuo to afford the crude Boc-intermediate as a whitesolid. The material was dissolved in 7 mL of DCM and treated with 4.0 mLof TFA. After two hours, the reaction solution was concentrated in vacuoto afford a pale yellow oil. The contents were partitioned between ethylacetate and diluted NaHCO₃ solution. The aqueous was extracted withethyl acetate, and the organics were combined. The organic was washedwith brine, separated, dried over Na₂SO₄, filtered, and concentrated invacuo. The residue was purified by chromatography (silica gel elutedwith 9:1 MeOH:DCM) to afford the pure2(R)-amino-3-(4-chlorophenyl)-N-(1-quinazolin-4-yl-azetidin-3-yl)-propionamideas a colorless oil (177 mg, 30%). ¹H NMR (CDCl₃, 400 MHz) δ 8.60 (s,1H), 8.02 (brs, 1H), 7.82 (d, J=8.4 Hz, 1H), 7.75 (d, J=8.8 Hz, 1H),7.72 (t, J=8.4 Hz, 1H), 7.40 (t, J=7.6 Hz, 1H), 7.29 (d, J=8.4 Hz, 1H),7.15 (d, J=8.4 Hz, 1H), 4.85 (m, 3H), 4.29 (dd, J=15.6, 5.6 Hz, 2H),3.64 (dd, J=8.8, 4.0 Hz, 1H), 3.22 (dd, J=13.6, 4.0 Hz, 1H), 2.78 (dd,J=14.0, 8.8 Hz, 1H). LCMS (APCI+) m/z 382 [M+H]⁺; Rt=0.76 min.

Example 89

Preparation of4-Amino-1-[4-(6-amino-5-cyclopropyl-pyrimidin-4-yl)-piperazin-1-yl]-2-(4-dichlorophenyl)-butan-1-onedihydrochloride

Step 1: To a stirred solution of 5-cyclopropyl-pyrimidine-4,6-diol (1.35g, 8.87 mmol) in DCE (35 mL) was added slowly POCl₃ (4.14 mL, 44.4 mmol)followed by DIEA (1.72 g, 13.3 mmol). The reaction mixture was heated toreflux for 2 days. After cooling, the solvent was evaporated in vacuo.The residue was partitioned between 5% NaHCO₃ and EtOAc. The organicphase was washed with brine, dried, and passed through a Silica gel padto give 4,6-dichloro-5-cyclopropyl-pyrimidine (1.30 g, 78%) as a yellowoil. ¹H NMR (CDCl₃, 400 MHz) δ (s, 1H), 1.65 (m, 1H), 1.03 (m, 2H), 1.58(m, 2H).

Step 2:4-(5-Cyclopropyl-6-chloro-pyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester was prepared by the procedures described in Example 40,Step 1, substituting 4-chloro-5-iodopyrimidine with5-cyclopropyl-4,6-dichloro-pyrimidine. ¹H NMR (CDCl₃, 400 MHz) δ 8.31(s, 1H), 3.64 (m, 4H), 3.54 (m, 4H), 1.73 (m, 1H), 1.46 (s, 9H), 1.14(m, 2H), 0.65 (m, 2H). LCMS (APCI+) m/z 339, 341 [M+H]⁺; Rt=2.38 min.

Step 3: A round bottom flask was charged with Pd(OAc)₂ (84 mg, 0.37mmol) and rac-BINAP (234 mg, 0.37 mmol) and purged with N₂. To the flaskwas added4-(5-cyclopropyl-6-chloro-pyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester (1.27 g, 3.75 mmol), benzophenone imine (815 mg, 4.50mmol), NaOBu^(t) (793 mg, 8.25 mmol) and toluene (24 mL). The mixturewas heated to 95° C. for 1 hour. After cooling to room temperature, thereaction was diluted with EtOAc, filtered through Celite, andconcentrated. The crude product was purified by column chromatography(hexanes:EtOAc, 2:1) to give4-[6-(Benzhydrylidene-amino)-5-cyclopropyl-pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester (1.62 g, 89%) as a colorless oil. ¹H NMR (CDCl₃,400 MHz) δ 8.38 (s, 1H), 7.41 (m, 10H), 3.40 (m, 4H), 3.32 (m, 4H), 1.46(s, 9H), 0.93 (m, 1H), 0.86 (m, 2H), 0.59 (m, 2H). LCMS (APCI+) m/z 484[M+H]⁺; Rt=3.94 min.

Step 4: To a stirred solution of4-[6-(Benzhydrylidene-amino)-5-cyclopropyl-pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester (1.60 g, 3.31 mmol) in MeOH (70 mL) was addedhydroxylamine hydrochloride (0.41 g, 6.0 mmol) and NaOAc (0.65 g, 7.9mmol). After stirring at room temperature overnight, the reactionmixture was partitioned between 0.1 N NaOH and DCM. The organic layerwas dried and concentrated. The residue was purified by column(DCM:MeOH, 20:1) to give4-(6-Amino-5-cyclopropyl-pyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester (1.00 g, 95%) as a colorless syrup. ¹H NMR (CDCl₃, 400MHz) δ 8.12 (s, 1H), 4.97 (s, 2H), 3.52 (m, 4H), 3.50 (m, 4H), 1.48 (s,9H), 1.44 (m, 1H), 1.02 (m, 2H), 0.64 (m, 2H). LCMS (APCI+) m/z 320[M+H]⁺; Rt=2.51 min.

Step 5: 5-Cyclopropyl-6-piperazin-1-yl-pyrimidin-4-ylaminedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-Chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with4-(6-Amino-5-cyclopropyl-pyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.18 (s, 1H), 4.12 (m, 4H),3.31 (m, 4H), 1.66 (m, 1H), 1.18 (m, 2H), 0.58 (m, 2H). LCMS (APCI+) m/z220 [M+H]⁺; Rt=0.74 min.

Step 6:4-Amino-1-[4-(6-amino-5-cyclopropyl-pyrimidin-4-yl)-piperazin-1-yl]-2-(4-chlorophenyl)-butan-1-onedihydrochloride was prepared by substituting5-piperazin-1-yl-1H-indazole with5-Cyclopropyl-6-piperazin-1-yl-pyrimidin-4-ylamine dihydrochloride andsubstituting (D)-Boc-4-chlorophenylalanine with4-tert-Butoxycarbonylamino-2-(4-chlorophenyl)-butyric acid in Example34, Step 2, then removing the Boc protecting group as described inExample 34, Step 3. ¹H NMR (CD₃OD, 400 MHz) δ 8.04 (s, 1H), 7.41 (d,J=8.4 Hz, 2H), 7.33 (d, J=8.4 Hz, 2H), 4.19 (m, 1H), 3.95 (m, 1H), 3.85(m, 2H), 3.61 (m, 3H), 3.22 (m, 2H), 2.95 (m, 1H), 2.82 (m, 1H), 2.31(m, 1H), 2.01 (m, 1H), 1.56 (m, 1H), 1.08 (m, 2H), 0.47 (m, 2H). LCMS(APCI+) m/z 415, 417 [M+H]⁺; Rt=1.87 min.

Example 90

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-furo[3,2-b]pyridin-7-yl-piperazin-1-yl)-propan-1-onedihydrochloride

Step 1: A solution of 3-hydroxy-2-bromopyridine (21.4 g, 123 mmol) inacetic anhydride (25 g, 245 mmol) was refluxed for 1 hour. Aftercooling, the mixture was poured into ice water, neutralized with Na₂CO₃and extracted with ether. The organic phase was dried and concentrated.The residue was subject to column chromatography, eluted by ether toafford Acetic acid 2-bromo-pyridin-3-yl ester (26.2 g, 99%). ¹H NMR(CDCl₃, 400 MHz) δ 8.28 (m, 1H), 7.46 (m, 1H), 7.30 (m, 1H), 2.38 (s,3H).

Step 2: To a solution of PdCl₂(PPh₃)₂ (1.4 g, 2.0 mmol) and CuI (0.4 g,2.1 mmol) in TEA (100 mL) and THF (200 mL) under nitrogen was added amixture of 2-bromo-3-acetoxy-pyridine (13.1 g, 60.6 mmol) andTMS-acetylene (7.0 g, 71 mmol) in THF (100 mL) in one portion. Themixture was stirred at room temperature for 1 hour. Then quenched withsaturated NaHCO₃ (50 mL) and MeOH (50 mL). The mixture was stirred at80° C. for 2 hours. After cooling, the mixture was extracted with ether.The organic phase was dried and concentrated. The residue was subject tocolumn chromatography to afford Furo[3,2-b]pyridine (1.54 g, 21%). ¹HNMR (CDCl₃ 400 MHz) δ 8.55 (m, 1H), 7.84 (m, 1H), 7.75 (m, 1H), 7.72 (m,1H), 6.99 (m, 1H).

Step 3: To a solution of Furo[3,2-b]pyridine (1.5 g, 13.0 mmol) in CHCl₃(30 mL) was added MCPBA (2.9 g, 17.0 mmol). The mixture was stirred atroom temperature for 16 hours. Then the mixture was filtered through aalumina (140 g, basic) and washed with ethyl acetate/hexane (1:1) andDCM/MeOH (20:1) to give Furo[3,2-b]pyridine 4-oxide (1.49 g, 88%). ¹HNMR (CDCl₃,400 MHz) δ 8.25 (m, 1H), 7.81 (m, 1H), 7.51 (m, 1H), 7.23 (m,1H).

Step 4: To a solution of Furo[3,2-b]pyridine 4-oxide in CHCl₃ (5 mL) wasadded POCl₃ (5 g, 33 mmol). The mixture was refluxed for 2 hours. Aftercooling, the mixture was quenched with ice water and neutralized withNaHCO₃. Extracted with CHCl₃ (3×100 mL). The organic phase was dried andconcentrated. The residue was subject to column chromatography to afford7-Chloro-furo[3,2-b]pyridine (0.74 g, 50%). ¹H NMR (CDCl₃, 400 MHz) δ8.46 (m, 1H), 7.92 (m, 1H), 7.28 (m, 1H), 7.04 (m, 1H).

Step 5: A mixture of 7-Chloro-furo[3,2-b]pyridine (0.73 g, 4.75 mmol)and piperazine (1.2 g, 14 mmol) in a sealed tube was heated to 130° C.for 4 hours. After cooling, the solid was dissolved in MeOH and DCM,concentrated and subject to column chromatography, eluted by DCM/MeOH(10:1-1:1) to give 7-piperazin-1-yl-furo[3,2-b]pyridine (0.21 g, 22%).MS (APCI+) [M+H]⁺ 204.

Step 6: To a solution of 7-piperazin-1-yl-furo[3,2-b]pyridine (0.21 g,1.03 mmol) and(2R)-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid (0.6g, 2.0 mmol) in DMF (10 mL) and TEA (2 mL) was added HOBT (0.3 g, 2.2mmol) and EDCI (0.42 g, 2.2 mmol). The mixture was stirred at roomtemperature for 4 hours. The solvent was removed and the residue wassubject to column chromatography to afford(2R)-[1-(4-Chlorobenzyl)-2-(4-furo[3,2-b]pyridin-7-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester (7 mg, 1.4%). MS (APCI+) [M+H]⁺ 486.

Step 7: To a solution of(2R)-[1-(4-Chlorobenzyl)-2-(4-furo[3,2-b]pyridin-7-yl-piperazin-1-yl)-2-oxo-ethyl]-carbamicacid tert-butyl ester (7 mg, 0.014 mmol) in DCM (4 mL) was added HCl indioxane (4M, 1 mL). The mixture was stirred at room temperature for 4hours. The solvent was removed to give(2R)-2-Amino-3-(4-chlorophenyl)-1-(4-furo[3,2-b]pyridin-7-yl-piperazin-1-yl)-propan-1-oneas HCl salt (6 mg, 99%). MS (APCI+) [M+H]⁺ 386.

Example 91

Preparation of(7S,2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(7-methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride and(7R,2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(7-methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: A solution of 2-Chloro-6-methyl-nicotinic acid (3.00 g, 17.5mmol) in ammonia in MeOH (7M, 60 mL) in a bomb was heated to 120° C.overnight. After cooling, the solvent was removed and the residue wasneutralized with 2N HCl. The precipitate was filtered, washed with waterand dried to afford 2-Amino-6-methyl-nicotinic acid (1.44 g, 54%). ¹HNMR (CD₃OD, 400 MHz) δ 8.21 (d, J=7.6 Hz, 1H), 6.60 (d, J=7.6 Hz, 1H),2.41 (s, 3H). MS (APCI+) [M+H]⁺ 153.

Step 2: A mixture of 2-Amino-6-methyl-nicotinic acid (1.44 g, 9.46 mmol)and formamide (8.0 g, 178 mmol) was stirred at 170° C. for 2 hours.After cooling, the mixture was quenched with water (4 mL). Theprecipitate was filtered, washed with water and dried to afford7-Methyl-pyrido[2,3-d]pyrimidin-4-ol (0.79 g, 51%). ¹H NMR (CDCl₃, 400MHz) δ 8.49 (d, J=8.4 Hz, 1H), 8.22 (s, 1H), 7.35 (d, J=8.0 Hz, 1H),2.75 (s, 3H). MS (APCI+) [M+H]⁺ 162.

Step 3: To a solution of 7-Methyl-pyrido[2,3-d]pyrimidin-4-ol (0.78 g,4.84 mmol) in DCE (30 mL) was added DIEA (1.0 mL, 1.19 mmol), followedby POCl₃ (2.4 mL, 26.1 mmol). The mixture was refluxed overnight. Aftercooling, the solvent was removed and the residue was dissolved in water(50 mL) and extracted with ethyl acetate (3×100 mL). The organic phasewas dried and concentrated. The residue was subject to columnchromatography, eluted by hexane/ethyl acetate (2:1) to give4-Chloro-7-methyl-pyrido[2,3-d]pyrimidine (0.66 g, 76%). ¹H NMR (CDCl₃,400 MHz) δ 9.22 (s, 1H), 8.49 (d, J=8.8 Hz, 1H), 7.57 (d, J=8.4 Hz, 1H),2.88 (s, 3H). MS (APCI+) [M+H]⁺ 180.

Step 4: The mixture of 4-Chloro-7-methyl-pyrido[2,3-d]pyrimidine (0.66g, 3.67 mmol) and 1-Boc-piperazine (0.75 g, 4.03 mmol) in DCE (40 mL)and TEA (5 mL) was refluxed for 1 hour. After cooling, the solvent wasremoved and the residue was subject to column chromatography, eluted byethyl acetate-DCM/MeOH (10:1) to give4-(7-Methyl-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester (1.2 g, 99%). ¹H NMR (CDCl₃, 400 MHz) δ 8.82 (s, 1H),8.11 (d, J=8.4 Hz, 1H), 7.28 (d, J=8.8 Hz, 1H), 3.79 (m, 4H), 3.65 (m,4H), 2.75 (s, 3H), 1.50 (s, 9H). MS (APCI+) [M+H]⁺ 330.

Step 5: A solution of4-(7-Methyl-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester (1.2 g, 3.64 mmol), PtO₂ (42 mg, 0.18 mmol) in MeOH (40mL) and TFA (2 mL) was stirred under H₂ (1 atm) at room temperature for4 hours. The catalyst was filtered and the solvent was removed. Theresidue was subject to column chromatography, eluted by DCM/MeOH (20:1)to give4-(7-Methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester as TFA salt (0.43 g, 27%). ¹H NMR (CDCl₃, 400 MHz)δ 9.75 (s, 1H), 7.97 (s, 1H), 3.60 (m, 10H), 2.60 (m, 2H), 2.00 (m, 1H),1.48 (s, 9H), 1.35 (d, J=6.4 Hz, 3H). MS (APCI+) [M+H]⁺ 334.

Step 6: To a solution of4-(7-Methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester TFA salt (0.19 g, 0.44 mmol) in DCM (20 mL) andTEA (1 mL) was added (S)-Mosher's acid chloride (0.14 g, 0.55 mmol). Themixture was stirred at room temperature for 20 minutes. The solvent wasremoved and the residue was subject to column chromatography, eluted byhexane/ethyl acetate (4:1-3:1-2:1). The first spot gave(7S)-4-[7-Methyl-8-(3,3,3-trifluoro-2-methoxy-2-phenyl-propionyl)-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester (84 mg, 34%). ¹H NMR (CDCl₃, 400 MHz) δ 8.27 (s,1H), 7.28 (m, 2H), 7.12 (m, 3H), 4.74 (m, 1H), 3.90 (s, 3H), 3.40 (m,4H), 3.08 (m, 4H), 2.32 (m, 1H), 2.20 (m, 1H), 1.94 (m, 1H), 1.49 (s,9H), 1.27 (m, 1H), 1.11 (d, J=6.4 Hz, 3H). MS (ESI+) [M+H]⁺ 550 Thesecond spot gave(7R)-4-[7-Methyl-8-(3,3,3-trifluoro-2-methoxy-2-phenyl-propionyl)-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester (85 mg, 35%). ¹H NMR (CDCl₃, 400 MHz) δ 8.20 (s,1H), 7.57 (m, 2H), 7.40 (m, 3H), 4.54 (m, 1H), 3.52 (s, 3H), 3.50 (m,4H), 3.31 (m, 4H), 2.55 (m, 1H), 2.42 (m, 1H), 1.91 (m, 1H), 1.48 (s,9H), 1.27 (m, 1H), 1.19 (d, J=6.4 Hz, 3H). MS (APCI+) [M+H]⁺ 550.

Step 7: To a solution of(7S)-4-[7-Methyl-8-(3,3,3-trifluoro-2-methoxy-2-phenyl-propionyl)-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester (84 mg, 0.153 mmol) in MeOH (5 mL) was added LiOH(3M, 4 mL). The mixture was stirred at room temperature for 4 days andthen neutralized with 2N HCl. The solvent was removed and the residuewas subject to column chromatography, eluted by ethyl acetate-DCM/MeOH(20:1) to afford(7S)-4-(7-Methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (32 mg, 63%). ¹H NMR (CDCl₃, 400 MHz) δ 8.11 (s,1H), 5.20 (s, 1H), 3.54 (m, 5H), 3.30 (m, 4H), 2.57 (m, 2H), 1.95 (m,1H), 1.48 (s, 9H), 1.40 (m, 1H), 1.27 (d, J=6.4 Hz, 3H). MS (APCI+)[M+H]⁺ 334.

Step 8: To a solution of(7S)-4-(7-Methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (32 mg, 0.096 mmol) in DCM (2 mL) was added HCl indioxane (4M, 2 mL). The mixture was stirred at room temperature for 3hours. The solvent was removed to give(7S)-7-Methyl-4-piperazin-1-yl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidineas HCl salt (22 mg, 99%). MS (APCI+) [M+H]⁺ 234.

Step 9: To a solution of(7S)-7-Methyl-4-piperazin-1-yl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidine(7.5 mg, 0.032 mmol) in DCM (4 mL) and DIEA (0.5 mL) was addedD-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid (10 mg,0.033 mmol) and HBTU (13 mg, 0.034 mmol). The mixture was stirred atroom temperature for 1 hour. The solvent was removed and the residue wassubject to column chromatography, eluted by hexane/ethyl acetate (2:1)to give(7S,2R)-{1-(4-Chlorobenzyl)-2-[4-(7-methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (16 mg, 96%). ¹H NMR (CDCl₃, 400 MHz) δ 8.10 (s,1H), 7.26 (d, J=6.8 Hz, 2H), 7.12 (d, J=8.0 Hz, 2H), 5.45 (d, J=8.4 Hz,1H), 5.10 (s, 1H), 4.84 (m, 1H), 3.60 (m, 4H), 3.30 (m, 2H), 3.00 (m,4H), 2.50 (m, 2H), 1.96 (m, 1H), 1.80 (m, 1H), 1.44 (m, 1H), 1.41 (s,9H), 1.27 (d, J=6.4 Hz, 3H). MS (APCI+) [M+H]⁺ 516.

Step 10: To a solution of(7S,2R)-{1-(4-Chlorobenzyl)-2-[4-(7-methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (16 mg, 0.031 mmol) in DCM (5 mL) was added HCl indioxane (4M, 2 mL). The mixture was stirred at room temperature for 4hours. The solvent was removed to give(7S,2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(7-methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (13 mg, 99%). MS (APCI+) [M+H]⁺ 416.(7R,2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(7-methyl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared in a similar manner from(7R)-4-[7-Methyl-8-(3,3,3-trifluoro-2-methoxy-2-phenyl-propionyl)-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester.

Example 92

Preparation of3-amino-2-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a solution of7-benzenesulfonyl-5-methyl-4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidinedihydrochloride (35 mg, 0.081 mmol),3-Boc-amino-2-(4-chlorophenyl)-propionic acid (27 mg, 0.089 mmol), andTEA (0.11 mL, 0.81 mmol) 1.2 mL DCM was added HBTU (34 mg, 0.089 mmol).The reaction mixture was stirred at room temperature 2 hours, afterwhich 0.15 mL 3M LiOH and 1.0 mL MeOH were added. The reaction mixturewas stirred at 35° C. for 3.5 hours, after which saturated NaHCO₃ wasadded. The mixture was extracted with DCM, and the combined extractswere dried (Na₂SO₄), filtered, and concentrated. The crude was purifiedon silica gel (flushed with 1:1 DCM:EtOAc, then gradient to 1:4DCM:EtOAc) to give3-Boc-amino-2-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-one,which was used in the next step.

Step 2: To a solution of3-Boc-amino-2-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onein 1 mL dioxane was added 1.5 mL 4M HCl/dioxane. The resultingsuspension was stirred at room temperature 7 hours, after which it wasconcentrated to dryness. The solids were then dissolved in minimal MeOH,and the product was triturated by the addition of ether. The solids wereisolated by filtration through a fritted funnel with nitrogen pressure,rinsed with ether, and dried in vacuo to afford3-amino-2-(4-chlorophenyl)-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (21 mg, 55%) as a beige powder. ¹H NMR (D₂O, 400 MHz) δ8.08 (s, 1H), 7.29 (d, J=8.4 Hz, 2H), 7.17 (d, J=8.4 Hz, 2H), 7.07 (s,1H), 4.30 (t, J=6.5 Hz, 1H), 4.03-3.94 (m, 1H), 3.82-3.73 (m, 1H),3.68-3.46 (m, 5H), 3.28 (dd, J=12.9, 7.4 Hz, 1H), 3.23 (dd, J=12.9, 5.7Hz, 1H), 3.04-2.94 (m, 1H), 2.13 (s, 3H). LCMS (APCI+) m/z 399 [M+H]⁺;Rt: 2.11 min.

Example 93

The Preparation of2-(4-Chlorophenyl)-3-dimethylamino-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onebis-hydrochloride salt

Step 1: The7-benzenesulfonyl-5-methyl-4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidinebis-hydrochloride (200 mg, 0.465 mmol) and3-tert-butoxycarbonylamino-2-(4-chlorophenyl)-propionic acid (146 mg,0.488 mmol) were dissolved/suspended in 2.0 mL of DMF at roomtemperature and treated with TEA (259 μL, 1.86 mmol). The HBTU (194 mg,0.511 mmol) was added in one sum, and the reaction was allowed to stirovernight at room temperature to completion. The reaction waspartitioned between ethyl acetate and diluted NaHCO₃ solution. Theaqueous was extracted with ethyl acetate, and the organics werecombined. The organic was washed with water, then brine, separated,dried over MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by chromatography (silica gel eluted with 70:30 ethylacetate:hexanes) to afford the pure[3-[4-(7-benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-(4-chlorophenyl)-3-oxo-propyl]-carbamicacid tert-butyl ester as colorless gel (295 mg, 99%). ¹H NMR (CDCl₃, 400MHz) δ 8.44 (s, 1H), 8.16 (d, J=8.4 Hz, 2H), 7.59 (t, J=7.2 Hz, 1H),7.49 (appt, J=7.6 Hz, 2H), 7.31 (m, 3H), 7.20 (d, J=8.0 Hz, 2H), 5.12(m, 1H), 4.09 (m, 1H), 3.92 (m, 1H), 3.60 (m, 1H), 3.56-3.46 (m, 4H),3.38 (d, J=10.4 Hz, 2H), 3.32 (m, 1H), 2.90 (m, 1H), 2.29 (s, 3H), 1.41(s, 9H). LCMS (APCI+) m/z 639 [M+H]⁺; Rt=3.70 min.

Step 2: The[3-[4-(7-benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-(4-chlorophenyl)-3-oxo-propyl]-carbamicacid tert-butyl ester (295 mg, 0.464 mmol) was dissolved in 2.3 mL of1,4-dioxane and treated with 4M HCl in 1,4-dioxane (2.3 mL, 9.29 mmol).The mixture was allowed to stir for 4 hours to completion affording ayellow precipitate. The suspension was diluted with diethyl ether andpoured into water. More diethyl ether was added, and the layers wereshaken. The ether wash was discarded, and the aqueous was treated withsaturated NaHCO₃ solution until basic to pH paper (about 10) to afford awhite precipitate. The aqueous was extracted with ethyl acetate, and theorganics were combined. The organic was washed with water, then brine,separated, dried over MgSO4, filtered, and concentrated in vacuo toafford the near-pure3-amino-1-[4-(7-benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-(4-chlorophenyl)-propan-1-oneas a colorless oil (194 mg, 77%). ¹H NMR (free-base, CDCl₃, 400 MHz) δ8.44 (s, 1H), 8.16 (d, J=8.4 Hz, 2H), 7.59 (t, J=7.2 Hz, 1H), 7.49(appt, J=7.6 Hz, 2H), 7.31 (m, 3H), 7.19 (d, J=8.0 Hz, 2H), 3.92 (m,1H), 3.85 (dd, J=8.4, 5.2 Hz, 1H), 3.61 (m, 1H), 3.56-3.50 (m, 4H), 3.43(m, 2H), 3.32 (m, 2H), 2.90 (m, 2H), 2.29 (s, 3H). LCMS (APCI+) m/z 539[M+H]⁺; Rt=2.40 min.

Step 3: The3-amino-1-[4-(7-benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-(4-chlorophenyl)-propan-1-one(75 mg, 0.139 mmol) was dissolved in 1.0 mL of 1,2-dichloroethane andtreated with 37% aqueous formaldehyde (31 μL, 0.417 mmol). The solutionwas allowed to stir at room temperature for 15 minutes before the sodiumtriacetoxyborohydride (118 mg, 0.557 mmol) was added in one sum. Thereaction was complete in one hour, and the contents were poured intodiluted NaHCO₃ solution. The aqueous was extracted with ethyl acetate,and the organics were combined. The organic was washed with water, thenbrine, separated, dried over MgSO₄, filtered, and concentrated in vacuo.The residue was purified by chromatography (silica gel eluted with 1%TEA in 9:1 ethyl acetate:methanol) to afford the pure1-[4-(7-benzenesulfonyl-5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-(4-chlorophenyl)-3-dimethylamino-propan-1-oneintermediate as a colorless oil. The oil was dissolved in 0.5 mL ofeach: THF, methanol, and water. The solution was treated with lithiumhydroxide-monohydrate (29 mg, 0.696 mmol) to afford an opaque solution,which stirred overnight to completion. The solution was partitionedbetween ethyl acetate and water, and the aqueous was extracted with moreethyl acetate. The combined organic was washed with water, then brine,separated, dried over Na₂SO₄, filtered, and concentrated in vacuo toafford the crude product as a yellow oil. The residue was purified bychromatography (silica gel eluted with 1% TEA in 4:1 ethylacetate:methanol) to afford the titled free base as a colorless oil. Thematerial was dissolved in a minimal amount of THF (>1 mL) and treatedwith 2.0M HCl in ether. The resulting white precipitate was filtered,washed with diethyl ether, and dried under reduced pressure to affordthe2-(4-chlorophenyl)-3-dimethylamino-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onebis-hydrochloride salt as a white solid (24 mg, 35%). ¹H NMR (Free-Base,CDCl₃, 400 MHz) δ 10.72 (brs, 1H), 8.34 (s, 1H), 7.30 (d, J=8.4 Hz, 2H),7.24 (d, J=8.4 Hz, 2H), 6.93 (s, 1H), 4.11 (m, 1H), 3.95 (m, 1H), 3.65(m, 2H), 3.55 (m, 3H), 3.33 (m, 2H), 3.05 (m, 1H), 2.48 (dd, J=12.4, 4.8Hz, 1H), 2.36 (s, 3H), 2.32 (s, 6H). LCMS (APCI+) m/z 427 [M+H]⁺;Rt=1.97 min.

Example 94

Preparation of2-(4-Chlorophenyl)-4-dimethylamino-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-butan-1-onebis-hydrochloride salt

The2-(4-chlorophenyl)-4-dimethylamino-1-[4-(5-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-butan-1-onebis-hydrochloride salt (21 mg, 30%) was prepared by procedures describedin Example 93, Steps 1-3 [substituting3-tert-butoxycarbonylamino-2-(4-chlorophenyl)-propionic acid with4-tert-butoxycarbonylamino-2-(4-chlorophenyl)-butyric acid in Step 1].¹H NMR (CD₃OD, 400 MHz) δ 8.29 (s, 1H), 7.41 (d, J=8.4 Hz, 2H), 7.36 (d,J=8.4 Hz, 2H), 7.25 (s, 1H), 4.25 (t, J=6.8 Hz, 1H), 3.97 (appd, J=8.8Hz, 2H), 3.85-3.72 (m, 4H), 3.63 (m, 1H), 3.32 (m, 1H), 3.17 (m, 1H),3.07 (m, 1H), 2.89 (s, 3H), 2.88 (s, 3H), 2.41 (m, 1H), 2.39 (s, 3H),2.10 (m, 1H). LCMS (APCI+) m/z 441 [M+H]⁺; Rt=1.87 min.

Example 95

Preparation ofN-(2-Amino-ethyl)-N-(4-chlorobenzyl)-4-pyridin-4-yl-benzamidebis-hydrochloride salt

Step 1: The (2-amino-ethyl)-carbamic acid tert-butyl ester (5.00 g, 31.2mmol) and 4-chloro-benzaldehyde (4.61 g, 32.77 mmol) were dissolved in60 mL of 1,2-dichloroethane at room temperature. The reaction mixturewas allowed to stir for 40 minutes prior to treatment with sodiumtriacetoxyborohydride (9.90 g, 46.8 mmol). The mixture was allowed tostir overnight to completion and quenched with a saturated NaHCO₃solution. The aqueous was extracted with DCM, separated, dried overMgSO₄, and concentrated in vacuo. The residue was purified bychromatography (silica gel eluted with hexanes/EtOAc plus 2% triethylamine) to afford the pure [2-(4-chlorobenzylamino)-ethyl]-carbamic acidtert-butyl ester as a viscous yellow oil (5.38 g, 61%). ¹H NMR (CDCl₃,400 MHz) δ 7.29 (d, J=8.0 Hz, 2H), 7.25 (d, J=8.0 Hz, 2H), 4.92 (brs,1H), 3.75 (s, 2H), 3.23 (appd, J=5.6 Hz, 2H), 2.73 (appt, J=6.0 Hz, 2H),1.44 (s, 9H), 1.36 (brs, 1H). LCMS (APCI+) m/z 285 [M+H]⁺; Rt=2.30 min.

Step 2: The 4-bromo-benzoic acid ethyl ester (1.00 g, 4.37 mmol),pyridine-4-boronic acid (537 mg, 4.37 mmol), andtetrakis(triphenylphosphine)palladium(0) (757 mg, 0.655 mmol) weredegassed under nitrogen. The solids were dissolved in 15 mL of1,4-dioxane and 2.6 mL of 2M sodium carbonate solution. The mixture washeated to 80° C. overnight completion and allowed to cool to roomtemperature. The contents were partitioned between ethyl acetate andwater, and the aqueous was extracted with ethyl acetate. The combinedorganic was washed with water, brine, dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was purified by chromatography(silica gel eluted with 1:1 hexanes:EtOAc, Rf=0.2) to afford the pure4-pyridin-4-yl-benzoic acid ethyl ester as a pale yellow solid (350 mg,35%). ¹H NMR (CDCl₃, 400 MHz) δ 8.71 (d, J=4.4 Hz, 2H), 8.16 (d, J=8.0Hz, 2H), 7.71 (d, J=8.0 Hz, 2H), 7.53 (d, J=4.4 Hz, 2H), 4.42 (q, J=7.2Hz, 2H), 1.43 (t, J=7.2 Hz, 3H). LCMS (APCI+) m/z 228 [M+H]⁺; Rt=2.81min.

Step 3: The 4-pyridin-4-yl-benzoic acid ethyl ester (430 mg, 1.89 mmol)was heated to 100° C. in 6.5 mL of 3M HCl solution overnight tocompletion. The mixture was cooled to room temperature and diluted withwater (dissolves precipitate). The solution was filtered andconcentrated in vacuo to afford the 4-pyridin-4-yl-benzoic acidhydrochloride salt as a pale yellow solid (380 mg, 85%). ¹H NMR(DMSO-d₆, 400 MHz) δ 8.93 (d, J=5.6 Hz, 2H), 8.27 (d, J=5.6 Hz, 2H),8.13 (d, J=8.0 Hz, 2H), 8.09 (d, J=8.0 Hz, 2H), —CO₂H or —NH⁺ notobserved. LCMS (APCI+) m/z 200 [M+H]⁺; Rt=1.05 min.

Step 4: The 4-pyridin-4-yl-benzoic acid hydrochloride salt (100 mg,0.502 mmol), [2-(4-chlorobenzylamino)-ethyl]-carbamic acid tert-butylester (143 mg, 0.502 mmol), HOBt (77 mg, 0.502 mmol), and EDCI (56 mg,0.552 mmol) were dissolved in 1.7 mL DMF, then treated with triethylamine (77 μL, 0.552 mmol) to room temperature. The reaction was allowedto stir overnight to completion and partitioned between ethyl acetateand diluted NaHCO₃ solution. The aqueous was extracted with ethylacetate, and the organics were combined. The organic was washed withwater, brine, dried over MgSO₄, filtered, and concentrated in vacuo. Theresidue was purified by chromatography (silica gel eluted withhexanes/ethyl acetate gradients plus 2% triethylamine) to afford the{2-[(4-chlorobenzyl)-(4-pyridin-4-yl-benzoyl)-amino]-ethyl}-carbamicacid tert-butyl ester as a colorless oil (184 mg, 93%). ¹H NMR (CDCl₃,400 MHz) δ 8.68 (d, J=4.8 Hz, 2H), 7.65 (m, 2H), 7.54 (d, J=8.4 Hz, 2H),7.48 (d, J=4.4 Hz, 2H), 7.34 (m, 2H), 7.13 (m, 2H), 4.91 (m, 1H), 4.59(brs, 2H), 3.62 (m, 2H), 3.44 (m, 2H), 1.46 (s, 9H). LCMS (APCI+) m/z466 [M+H]⁺; Rt=3.19 min.

Step 5: The{2-[(4-chlorobenzyl)-(4-pyridin-4-yl-benzoyl)-amino]-ethyl}-carbamicacid tert-butyl ester (184 mg, 0.395 mmol) was dissolved in 2.0 mL of1,4-dioxane and treated with 2.0 mL of 4M HCl in 1,4-dioxane. Thereaction was allowed to stir for one hour to completion and diluted withdiethyl ether. The product was isolated by vacuum filtration to affordthe N-(2-amino-ethyl)-N-(4-chlorobenzyl)-4-pyridin-4-yl-benzamidebis-hydrochloride salt as a fine pale-yellow powder (166 mg, 96%). ¹HNMR (DMSO-d₆, 400 MHz) δ 8.91 (d, J=6.0 Hz, 2H), 8.26 (d, J=6.0 Hz, 2H),8.13 (brs, 3H), 8.05 (d, J=8.0 Hz, 2H), 7.76 (d, J=7.6 Hz, 2H), 7.45(brs, 1H), 7.44 (d, J=8.0 Hz, 2H), 7.26 (d, J=7.6 Hz, 2H), 4.57 (brs,2H), 3.61 (m, 2H), 3.08 (m, 2H). LCMS (APCI+) m/z 366 [M+H]⁺; Rt=2.13min.

Example 96

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: A mixture of 2-aminonicotinic acid (7.00 g, 50.7 mmol) andformamide (22.8 g, 506 mmol) was heated to 167° C. for 2.5 hours. Aftercooling, the solid was recrystallized from hot water (100 mL) to affordthe pure product (4.80 g, 64%). ¹H NMR (CDCl₃, 400 MHz) δ 8.94 (m, 1H),8.64 (d, 1H, J=7.6 Hz), 8.30 (s, 1H), 7.59 (m, 1H). MS (APCI+) [M+H]⁺148.

Step 2: The solution of 4-hydroxypyrido[2,3-d]pyrimidine (2.00 g, 13.6mmol) in POCl₃ (40 mL) was refluxed for 2 hours. After cooling, theexcess POCl₃ was removed under vacuum. The residue was quenched withsaturated NaHCO₃ and extracted with ethyl acetate (3×100 mL). Theorganic phase was dried and concentrated. The residue was subject tocolumn chromatography, eluted by hexane/ethyl acetate (1:1) to give4-chloropyrido[2,3-d]pyrimidine (0.72 g, 32%). ¹H NMR (CDCl₃, 400 MHz) δ9.35 (m, 1H), 9.30 (s, 1H), 8.66 (m, 1H), 7.73 (m, 1H). MS (APCI+)[M+H]⁺ 166.

Step 3: To a solution of 4-chloropyrido[2,3-d]pyrimidine (0.72 g, 4.4mmol) and 1-Boc piperazine (0.84 g, 4.5 mmol) in DCE (10 mL) and IPA (10mL) was added triethylamine (4 mL). The mixture was refluxed for 4hours. After cooling, the solvent was removed. The residue was subjectto column chromatography by ethyl acetate to give4-pyrido[2,3-d]pyrimidin-4-yl-piperazine-1-carboxylic acid tert-butylester (1.26 g, 92%). ¹H NMR (CDCl₃, 400 MHz) δ 9.07 (dd, J=4.30 Hz,J=1.98 Hz, 1H), 8.88 (s, 1H), 8.25 (dd, J=7.99 Hz, J=1.98 Hz, 1H), 7.41(dd, J=8.18 Hz, J=3.74 Hz, 1H), 3.82 (m, 4H), 3.65 (m, 4H), 1.50 (s,9H). MS (APCI+) [M+H]⁺ 316.

Step 4: To a solution of4-pyrido[2,3-d]pyrimidin-4-yl-piperazine-1-carboxylic acid tert-butylester (0.24 g, 0.76 mmol) in MeOH (10 mL) and TFA (1 mL) was added PtO₂(10 mg). The mixture was stirred under H₂ (1 atm) at room temperatureovernight. The catalyst was filtered off and the solvent was removed.The residue was subject to column chromatography, eluted by DCM/MeOH(20:1) to afford4-(5,6,7,8-Tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (0.15 g, 62%). ¹H NMR (CDCl₃, 400 MHz) δ 8.10 (s,1H), 5.92 (s, 1H), 3.50 (m, 4H), 3.40 (m, 2H), 3.25 (m, 4H), 2.55 (m,2H), 1.85 (m, 2H), 1.48 (s, 9H). MS (APCI+) [M+H]⁺ 320.

Step 5: To a solution of4-(5,6,7,8-Tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (0.88 g, 2.75 mmol) in DCM (10 mL) was added HClin dioxane (4M, 5 mL). The mixture was stirred at RT for 4 hours. Thesolvent was removed to give4-piperazin-1-yl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidine as HCl salt(0.80 g, 99%). MS (APCI+) [M+H]⁺ 220.

Step 6: To a solution of4-piperazin-1-yl-5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidinedihydrochloride (20 mg, 0.068 mmol) and(2R)-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid (21mg, 0.070 mmol) in DCM (5 mL) and TEA (1 mL) was added HBTU (30 mg,0.079 mmol). The mixture was stirred at room temperature for 4 hours.The solvent was removed and the residue was subject to columnchromatography, eluted with ethyl acetate/DCM/MeOH (20:1) to give(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester (18 mg, 53%). ¹H NMR (CDCl₃, 400 MHz) δ 8.10 (s,1H), 7.25 (m, 3H), 7.12 (m, 2H), 5.42 (d, J=8.8 Hz, 1H), 5.14 (s, 1H),4.84 (m, 1H), 3.67 (m, 3H), 3.50 (m, 1H), 3.40 (m, 2H), 3.23 (m, 5H),2.95 (m, 4H), 2.50 (m, 2H), 1.86 (m, 2H), 1.67 (s, 1H), 1.42 (s, 9H). MS(APCI+) [M+H]⁺ 502.

Step 7: To a solution of(2R)-{1-(4-Chlorobenzyl)-2-oxo-2-[4-(5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-ethyl}-carbamicacid tert-butyl ester (18 mg, 0.036 mmol) in DCM (4 mL) was added HCl indioxane (4M, 2 mL). The mixture was stirred at room temperature for 4hours. The solvent was removed to give(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5,6,7,8-tetrahydro-pyrido[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (14 mg, 99%). MS (APCI+) [M+H]⁺ 401.

Example 97

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a solution of formamide HCl salt (21.6 g, 268 mmol) in MeOH(300 mL) was added NaOMe (25%, in MeOH, 120 mL, 555 mmol). The mixturewas stirred at room temperature for 1 hour. Then triethyl1,1,2-ethanetricarboxylate (64.4 g, 60 mL, 262 mmol) in MeOH (90 mL) wasadded slowly. After addition, the mixture was stirred at roomtemperature for 20 hours. The solvent was removed and the residue wasdissolved in ice water (200 mL) and neutralized with 2N HCl (140 mL)until pH=1-2. The solid formed was filtered, washed with water (50 mL)and dried under vacuum to afford (4,6-Dihydroxy-pyrimidin-5-yl)-aceticacid methyl ester (45 g, 93%). ¹H NMR (CDCl₃, 400 MHz) δ 8.03 (s, 1H),3.67 (s, 3H), 3.42 (s, 2H). MS (APCI+) [M+H]⁺ 185.

Step 2: To a solution of (4,6-Dihydroxy-pyrimidin-5-yl)-acetic acidmethyl ester (45 g, 244 mmol) in DCE (800 mL) was added DIEA (72 mL, 413mmol), followed by POCl₃ (80 mL, 874 mmol) slowly. After addition, themixture was stirred at room temperature for 2 hours and then refluxedovernight. After cooling, the solvent was removed and the residue wasdissolved in ice water (400 mL), neutralized with 10N NaOH until pH 6.Extracted with ethyl acetate (3×500 mL). The organic phase was dried andconcentrated. The residue was subject to column chromatography, elutedby hexane/ethyl acetate (4:1) to afford(4,6-Dichloro-pyrimidin-5-yl)-acetic acid methyl ester (38 g, 70%). ¹HNMR (CDCl₃, 400 MHz) δ 8.73 (s, 1H), 4.00 (s, 2H), 3.77 (s, 3H). MS(APCI+) [M+H]⁺ 222.

Step 3: To a solution of (4,6-Dichloro-pyrimidin-5-yl)-acetic acidmethyl ester (0.52 g, 2.35 mmol) in ether (40 mL) at −78° C. was addedDIBAL-H (1.5M, 4 mL, 6.05 mmol) dropwise. After addition, the mixturewas allowed to warm up to room temperature and stirred for 3 hours. Thenquenched with 2N HCl (10 mL) at −78° C. Extracted with ethyl acetate(3×50 mL). The organic phase was dried and concentrated to give fairlypure 2-(4,6-Dichloro-pyrimidin-5-yl)-ethanol (0.44 g, 97%). ¹H NMR(CDCl₃, 400 MHz) δ 8.66 (s, 1H), 3.96 (t, J=6.8 Hz, 2H), 3.23 (t, J=6.8Hz, 2H). 1.65 (s, 1H). MS (APCI+) [M+H]⁺ 194.

Step 4: To a solution of 2-(4,6-Dichloro-pyrimidin-5-yl)-ethanol (0.46g, 2.4 mmol) in DCM (40 mL) and was added MsCl (0.50 g, 4.36 mmol),triethylamine (1 mL) and cat. amount of DMAP. After stirring at roomtemperature overnight, the solvent was removed and the residue wassubject to column chromatography, eluted by hexane/ethyl acetate (4:1)to afford methanesulfonic acid 2-(4,6-dichloro-pyrimidin-5-yl)-ethylester (0.52 g, 80%). ¹H NMR (CDCl₃, 400 MHz) δ 8.71 (s, 1H), 4.50 (t,J=6.4 Hz, 2H), 3.42 (t, J=6.8 Hz, 2H), 3.02 (s, 3H).

Step 5: A solution of methanesulfonic acid2-(4,6-dichloro-pyrimidin-5-yl)-ethyl ester (0.50 g, 1.84 mmol) and4-methoxybenzylamine (0.60 g, 4.37 mmol) in DCE (30 mL) and TEA (4 mL)was refluxed overnight. After cooling, the solvent was removed and theresidue was subject to column chromatography, eluted by hexane/ethylacetate (4:1) to give4-Chloro-7-(4-methoxy-benzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine(0.36 g, 71%). ¹H NMR (CDCl₃, 400 MHz) δ 8.35 (s, 1H), 7.26 (dd, J=2.8Hz, J=1.6 Hz, 2H), 6.88 (d, J=4.4 Hz, 2H), 5.33 (s, 1H), 4.62 (d, J=5.6Hz, 2H), 3.81 (s, 3H), 3.73 (t, J=7.2 Hz, 2H), 3.04 (t, J=6.8 Hz, 2H).MS (APCI+) [M+H]⁺ 276.

Step 6: A solution of4-Chloro-7-(4-methoxy-benzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine(0.36 g, 1.31 mmol), 1-Boc-piperazine (1.0 g, 5.37 mmol) and tBuOK (0.18g, 1.60 mmol) in NMP (20 mL) was heated to 128° C. for 20 hours. Aftercooling, the mixture was diluted with ethyl acetate (500 mL) and washedwith water (5×150 mL). The organic phase was dried and concentrated. Theresidue was subject to column chromatography, eluted by hexane/ethylacetate (1:1) to give4-[7-(4-Methoxy-benzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester (0.32 g, 57%). ¹H NMR (CDCl₃, 400 MHz) δ 8.17 (s,1H), 7.20 (d, J=8.4 Hz, 2H), 6.84 (d, J=8.4 Hz, 2H), 4.50 (s, 2H), 3.79(s, 3H), 3.59 (m, 4H), 3.48 (m, 4H), 3.35 (m, 2H), 3.30 (m, 2H), 1.49(s, 9H). MS (APCI+) [M+H]⁺ 426.

Step 7: A solution of4-[7-(4-Methoxy-benzyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl]-piperazine-1-carboxylicacid tert-butyl ester (0.32 g, 0.74 mmol) in TFA (20 mL) was stirred at65° C. for 20 hours. After cooling, the TFA was evaporated under vacuumto afford 4-piperazin-1-yl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidinebis-trifluoroacetate (0.15 g, 99%). MS (APCI+) [M+H]⁺ 206.

Step 8: To a solution of4-piperazin-1-yl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidinebis-trifluoroacetate (20 mg, 0.097 mmol) in DCM (10 mL) and TEA (2 mL)were added (2R)-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionicacid (30 mg, 0.10 mmol) and HBTU (30 mg, 0.079 mmol). The mixture wasstirred at room temperature for 2 hours. The solvent was removed and theresidue was subject to column chromatography to afford(2R)-{1-(4-Chlorobenzyl)-2-[4-(6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (19 mg, 40%). ¹H NMR (CDCl₃, 400 MHz) δ 8.03 (s,1H), 7.25 (m, 2H), 7.12 (m, 2H), 5.40 (m, 1H), 4.83 (m, 2H), 3.50 (m,10H), 3.20 (m, 5H), 2.96 (m, 2H), 2.28 (m, 3H), 1.42 (s, 9H). MS (APCI+)[M+H]⁺ 488.

Step 9: To a solution of(2R)-{1-(4-Chlorobenzyl)-2-[4-(6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (19 mg, 0.039 mmol) in DCM (4 mL) was added HCl indioxane (4M, 2 mL). The mixture was stirred at room temperature for 6hours. The solvent was removed to afford(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (15 mg, 99%). MS (APCI+) [M+H]⁺ 388.

Example 98

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a solution of KH (30%, 13.3 g, 125 mmol) in THF (200 mL) at0° C. was added (4,6-Dichloro-pyrimidin-5-yl)-acetic acid methyl ester(20.0 g, 90.0 mmol) and MeI (290.0 g, 8.8 mL, 141.0 mmol) in THF (200mL) slowly. After addition, the mixture was stirred at room temperaturefor 30 min, and then heated to reflux for 1 hour. After cooling to 0°C., the mixture was quenched with saturated aqueous NH₄Cl. The organicphase was separated and the aqueous phase was extracted with ethylacetate (3×200 mL). The organic phase was combined and dried. Afterremoval of the solvent, the residue was subject to columnchromatography, eluted by hexane/ethyl acetate (5:1) to give2-(4,6-Dichloro-pyrimidin-5-yl)-propionic acid methyl ester (17.6 g,83%). ¹H NMR (CDCl₃, 400 MHz) δ 8.69 (s, 1H), 4.39 (dd, J=14.4 Hz, J=7.2Hz, 1H), 3.73 (s, 3H), 1.57 (d, J=7.2 Hz, 3H).

Step 2: To a solution of 2-(4,6-Dichloro-pyrimidin-5-yl)-propionic acidmethyl ester (0.5 g, 2.13 mmol) in ether (40 mL) at −78° C. was addedDIBAL-H (1.5M, 4 mL, 6.0 mmol) dropwise. The mixture was allowed to warmup to room temperature and stirred for 3 hours. Then quenched with 2NHCl (10 mL) at −78° C. The aqueous phase was extracted with ethylacetate (3×50 mL). The organic phase was dried and concentrated. Theresidue was subject to column chromatography, eluted by hexane/ethylacetate (4:1) to give 2-(4,6-Dichloro-pyrimidin-5-yl)-propan-1-ol (0.40g, 91%). ¹H NMR (CDCl₃, 400 MHz) δ 8.62 (s, 1H), 4.16 (m, 1H), 3.90 (m,2H), 1.93 (s, 1H), 1.40 (d, J=7.2 Hz, 3H).

Step 3: To a solution of 2-(4,6-Dichloro-pyrimidin-5-yl)-propan-1-ol(0.40 g, 1.93 mmol) in DCM (40 mL) were added MsCl (0.50 g, 4.36 mmol),TEA (1 mL) and catalytic amount of DMAP. The mixture was stirred at roomtemperature for 2 hours. The solvent was removed and the residue wassubject to column chromatography, eluted by hexane/ethyl acetate (4:1)to give methanesulfonic acid 2-(4,6-dichloro-pyrimidin-5-yl)-propylester (0.54 g, 98%). ¹H NMR (CDCl₃, 400 MHz) δ 8.68 (s, 1H), 4.74 (m,1H), 4.55 (m, 1H), 4.14 (m, 1H), 2.99 (s, 3H), 1.49 (dd, J=7.2 Hz, J=1.2Hz, 3H).

Step 4: To a solution of methanesulfonic acid2-(4,6-dichloro-pyrimidin-5-yl)-propyl ester (0.54 g, 1.89 mmol) in DCM(30 mL) and TEA (4 mL) was added 4-methoxybenzyl amine (0.80 g, 5.83mmol). The mixture was refluxed overnight. After cooling, the solventwas removed and the residue was subject to column chromatography, elutedby hexane/ethyl acetate (4:1) to give4-Chloro-7-(4-methoxy-benzyl)-5-methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine(0.55 g, 99%). ¹H NMR (CDCl₃, 400 MHz) δ 8.24 (s, 1H), 7.17 (d, J=7.6Hz, 2H), 6.86 (d, J=7.6 Hz, 2H), 4.55 (s, 2H), 3.80 (s, 3H), 3.66 (m,1H), 3.37 (m, 1H), 3.07 (m, 1H), 1.31 (dd, J=7.2 Hz, J=1.2 Hz, 3H). MS(APCI+) [M+H]⁺ 291.

Step 5: To a solution of4-Chloro-7-(4-methoxy-benzyl)-5-methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine(0.55 g, 1.88 mmol) in NMP (20 mL) were added 1-Boc-piperazine (1.0 g,5.40 mmol) and ^(t)BuOK (0.21 g, 1.88 mmol). The mixture was heated to128° C. for 30 hours. After cooling, the mixture was diluted by ethylacetate (500 mL) and washed with water (5×150 mL). The organic phase wasdried and concentrated. The residue was subject to columnchromatography, eluted by hexane/ethyl acetate (1:1) to give4-(5-Methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (0.30 g, 36%). ¹H NMR (CDCl₃, 400 MHz) δ 8.19 (s,1H), 7.18 (d, J=8.4 Hz, 2H), 6.84 (d, J=8.4 Hz, 2H), 4.52 (dd, J=27.6Hz, J=14.8 Hz, 2H), 3.79 (s, 3H), 3.58 (m, 12H), 3.34 (m 1H), 2.95 (m,1H), 1.48 (s, 9H), 1.15 (d, J=6.8 Hz, 3H). MS (APCI+) [M+H]⁺ 440.

Step 6: A solution of4-(5-Methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (0.30 g, 0.68 mmol) in TFA (20 mL) was heated to65° C. overnight. After cooling, the excess TFA was evaporated undervacuum to give5-Methyl-4-piperazin-1-yl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine as TFAsalt (0.15 g, 99%). MS (APCI+) [M+H]⁺ 220.

Step 7: To a solution of5-Methyl-4-piperazin-1-yl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (18mg, 0.082 mmol) in DCM (10 mL) and TEA (2 mL) were added(2R)-2-tert-Butoxycarbonylamino-3-(4-chlorophenyl)-propionic acid (25mg, 0.082 mmol) and HBTU (31 mg, 0.082 mmol). The mixture was stirred atroom temperature for 2 hours. The solvent was removed and the residuewas subject to column chromatography, eluted by ethyl acetate-DCM/MeOH(30:1) to give(2R)-{1-(4-Chlorobenzyl)-2-[4-(5-methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (32 mg, 78%). ¹H NMR (CDCl₃, 400 MHz) δ 8.06 (s,1H), 7.26 (d, J=6.0 Hz, 2 hours), 7.14 (d, J=8.0 Hz, 2H), 5.39 (d, J=8.0Hz, 1H), 4.82 (m, 2H), 3.45 (m, 10H), 3.18 (m, 4H), 3.00 (m, 2H), 1.83(m, 4H), 1.42 (s, 9H), 1.19 (m, 3H). MS (APCI+) [M+H]⁺ 502.

Step 8: To a solution of(2R)-{1-(4-Chlorobenzyl)-2-[4-(5-methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester (32 mg, 0.064 mmol) in DCM/MeOH (5:1, 6 mL) wasadded HCl in dioxane (4M, 2 mL). The mixture was stirred at roomtemperature for 6 hours. The solvent was removed to afford(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(5-methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride (26 mg, 99%). MS (APCI+) [M+H]⁺ 402.

Example 99

Preparation of(2R)—N-{4-[4-(2-amino-3-phenyl-propionyl)-piperazin-1-yl]-quinazolin-6-yl}-methanesulfonamidedihydrochloride

Step 1: To a solution of 4-chloro-6-nitroquinazoline (prepared accordingto the literature: Alexander J. Bridges et al. J. Med. Chem. 1996, 39,267-276, and references therein; 12 g, 57.5 mmol) and DIEA (10 mL, 57.5mmol) in 230 mL IPA was added Boc-piperazine (10.7 g, 57.5 mmol). Thereaction mixture was heated to 60° C. and stirred for 13 hours, afterwhich it was cooled to room temperature and concentrated by rotaryevaporation. The residue was dissolved in dichloromethane (DCM) andwashed with 1N NaOH. The organic layer was dried (Na₂SO₄), filtered, andconcentrated by rotary evaporation. The resulting oil was purified onsilica gel (1:1 to 1:4 DCM:EtOAc gradient) to furnish4-(6-nitro-quinazolin-4-yl)-1-Boc-piperazine as a pale yellow oil (18.3g, 89%). LCMS (APCI+) m/z 360 [M+H]⁺. HPLC Rt 3.06 min.

Step 2: To a suspension of Pd/C (5% w/w, 800 mg, 0.38 mmol) in 100 mL2-methoxyethanol (degassed with nitrogen prior to use) was added asolution of 4-(6-nitro-quinazolin-4-yl)-1-Boc-piperazine (4.0 g, 11.1mmol) in 10 mL 2-methoxyethanol. A balloon of H₂ was bubbled through thereaction mixture, and the reaction mixture was stirred at roomtemperature under an atmosphere of H₂ for 13 hours. Celite was thenadded, and the reaction mixture was filtered through a pad of celite andrinsed with MeOH. The filtrate was concentrated, and the resulting oilwas filtered through a short plug of silica gel with EtOAc. Theresulting filtrate was concentrated to give4-(6-amino-quinazolin-4-yl)-1-Boc-piperazine (3.47 g, 95%). LCMS (APCI+)m/z 330 [M+H]⁺. HPLC Rt 2.31 min.

Step 3: To a 0° C. solution of (6-aminoquinazolin-4-yl)1-Boc-piperazine(1.0 g, 3.04 mmol), triethylamine (1.7 mL, 12.2 mmol) and DMAP (93 mg,0.76 mmol) in 20 mL DCM was added dropwise by addition funnel a solutionof methanesulfonyl chloride (0.59 mL, 7.6 mmol) in 6 mL DCM. Thereaction mixture was stirred 5 minutes, warmed to room temperature, andstirred an additional 1.5 hours, after which the reaction mixture wascooled to 0° C., and NaOMe (5.4M in MeOH, 5.6 mL, 30.4 mmol) was addedslowly by syringe. The reaction mixture was stirred 10 minutes, warmedto room temperature, and stirred another 2 hours, after which saturatedNH₄Cl was added. The reaction mixture was extracted with DCM, and thecombined extracts were washed with brine, dried (Na₂SO₄), filtered, andconcentrated. The resulting residue was purified on silica gel (20:1DCM:MeOH) to giveN-(4-Boc-piperazin-1-yl-quinazolin-6-yl)-methanesulfonamide (1.05 g,85%) as a beige powder. LCMS (APCI+) m/z 408 [M+H]⁺. HPLC Rt 2.64 min.

Step 4: To a solution ofN-(4-Boc-piperazin-1-yl-quinazolin-6-yl)-methanesulfonamide (1.05 g,2.58 mmol) in 15 mL dioxane was added 10 mL 4M HCl/dioxane. Theresulting suspension was stirred at room temperature 17 hours, afterwhich it was diluted with ether, and the solids were isolated byfiltration through a fritted funnel with nitrogen pressure, rinsed withether, and dried in vacuo to furnishN-(4-piperazin-1-yl-quinazolin-6-yl)-methanesulfonamide dihydrochloride(969 mg, 99%) as a white powder. ¹H NMR (CD₃OD, 400 MHz) δ 8.80 (s, 1H),8.12 (s, 1H), 7.87-7.84 (m, 2H), 4.50 (dd, J=5.2, 5.2 Hz, 4H), 3.57 (dd,J=5.2, 5.2 Hz, 4H), 3.13 (s, 3H). LCMS (APCI+) m/z 308 [M+H]⁺. HPLC Rt1.55 min.

Step 5: To a Jones tube containing PS-CDI (Argonaut, 1.04 mmol/g, 56 mg,2.0 equiv) suspended in a solution ofN-(4-piperazin-1-yl-qyinazolin-6-yl)-mathanesulfonamide dihydrochloride(11 mg, 0.029 mmol, 1.0 equiv) and DIEA (25 μL, 0.15 mmol, 5.0 equiv) in1.6 mL 9:1 CHCl₃:THF were added successively HOBt.H₂O (6 mg, 0.038 mmol,1.3 equiv) and (D)-Boc-phenylalanine (8 mg, 0.032 mmol, 1.1 equiv.). Thereaction mixture was shaken for 15 hours at room temperature, afterwhich Si-trisamine (Silicycle, 1.21 mmol/g, 48 mg, 2.0 equiv) was added.The reaction mixture was shaken an additional 1 hour, after which it wasvacuum filtered, the resins rinsed with CHCl₃, and the filtrateconcentrated by rotary evaporation. The crude was purified on silica gel(19:1 DCM:MeOH) to afford(2R)—N-{4-[4-(2-Boc-amino-3-phenyl-propionyl)-piperazin-1-yl]-quinazolin-6-yl}-methanesulfonamideas a clear, colorless residue.

Step 6: To a solution of(2R)—N-{4-[4-(2-Boc-amino-3-phenyl-propionyl)-piperazin-1-yl]-quinazolin-6-yl}-methanesulfonamidein 1.0 mL dioxane was added 1.2 mL 4M HCl/dioxane. The resultingsuspension was stirred at room temperature another 13 hours, after whichit was concentrated to dryness. The resulting solids were dissolved inminimal MeOH, and the product was triturated by the addition of ether.The resulting suspension was diluted with ether, and the solids wereisolated by filtration through a fritted funnel with nitrogen pressure,rinsed with ether, and dried further in vacuo to afford(2R)—N-{4-[4-(2-amino-3-phenyl-propionyl)-piperazin-1-yl]-quinazolin-6-yl}-methanesulfonamidedihydrochloride (13 mg, 84%) as a yellow powder. LCMS (APCI+) m/z 455[M+H]⁺. HPLC Rt 2.18 min.

Example 100

Preparation of4-Amino-2-(2-chlorophenyl)-1-[4-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazin-1-yl]-butan-1-onebis-hydrochloride salt

Step 1: The 1H-Pyrazolo[3,4-d]pyrimidin-4-ol (5.00 g, 36.73 mmol) wasdissolved in 68.5 mL of phosphorous oxychloride and 9.31 mL ofN,N-dimethyl aniline (73.47 mmol). This mixture was heated to reflux(120 C) for 90 minutes to completion affording a dark red solution. Themixture was concentrated in vacuo and cooled to 0° C. in an ice bath.The residue was poured into ice water and stirred for three minutes. Theacidic melt was extracted with ether, and the organics were combined.The organic was washed with cold water, cold half saturated NaHCO₃solution, brine, separated, dried over MgSO₄, filtered, and concentratedin vacuo to afford the 4-chloro-1H-pyrazolo[3,4-d]pyrimidine as a lightyellow powder (2.30 g, 41%). ¹H NMR (DMSO-d₆, 400 MHz) δ 8.84 (s, 1H),8.46 (s, 1H), NH not observed.

Step 2: The 4-chloro-1H-pyrazolo[3,4-d]pyrimidine (500 mg, 3.24 mmol)and piperazine-1-carboxylic acid tert-butyl ester (603 mg, 3.24 mmol)were dissolved in 11.0 mL of NMP then treated with diisopropylethylamine (845 μL, 4.85 mmol). The yellow solution was heated to 80 C.overnight to completion and was allowed to cool to room temperature. Thesolution was diluted with ethyl acetate, poured into diluted NaHCO₃solution, and extracted with ethyl acetate. The combined organic waswashed with water, brine, separated, dried over MgSO₄, filtered, andconcentrated in vacuo to afford the crude material as a tan solid. Thematerial was triturated with DCM/hexanes to afford the4-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazine-1-carboxylic acidtert-butyl ester as a cream-colored solid (824 mg, 84%). ¹H NMR (CDCl₃,400 MHz) δ 8.48 (s, 1H), 8.06 (s, 1H), 4.05 (m, 4H), 3.67 (m, 4H). LCMS(APCI+) m/z 305 [M+H]⁺; Rt=2.14 min.

Step 3: The 4-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester (600 mg, 1.97 mmol) was dissolved in 4 mL of1,4-dioxane and treated with 10 mL of 4M HCl in 1,4-dioxane at roomtemperature. The solution was allowed to stir for two hours to afford alight-yellow suspension of product. The solvent was diluted with diethylether, stirred for ten minutes, and filtered. The pad of product waswashed with diethyl ether and allowed to dry under a stream of nitrogento give the 4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidinebis-hydrochloride salt as a light-yellow solid (539 mg, 99%). ¹H NMR(D₆O, 400 MHz) δ 8.59 (s, 1H), 8.42 (s, 1H), 4.29 (appt, J=5.6 Hz, 4H),3.42 (appt, J=5.6 Hz, 4H). LCMS (APCI+) m/z 205 [M+H]⁺; Rt=0.34 min.

Step 4: The 4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidinebis-hydrochloride salt (60 mg, 0.22 mmol), HOBt (29 mg, 0.22 mmol), EDCI(46 mg, 0.24 mmol), and4-tert-butoxycarbonylamino-2-(2-chlorophenyl)-butyric acid [prepared byprocedures described in Example 61 (68 mg. 0.22 mmol)] weresuspended/dissolved in 1.5 mL of DMF then treated with triethylamine(121 L, 0.87 mmol). The mixture was allowed to stir for four hours tocompletion then partitioned between ethyl acetate and diluted NaHCO₃solution. The aqueous was extracted with ethyl acetate, and the organicswere combined. The organic was washed with water, brine, separated,dried over MgSO₄, filtered, and concentrated in vacuo. The residue wasdissolved in 1 mL of 1,4-dioxane and treated with 1 mL of 4M HCl in1,4-dioxane. The solution stirred overnight to completion at roomtemperature and diluted with diethyl ether to afford a precipitate. Thismaterial was broken up to afford a suspended granular solid which wasstirred for 30 minutes. The suspension was filtered, washed with diethylether, and dried over a stream of nitrogen to afford the4-amino-2-(2-chlorophenyl)-1-[4-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazin-1-yl]-butan-1-onebis-hydrochloride salt as a tan solid (69.2 mg, 67%). ¹H NMR (D₆O, 400MHz) δ 8.52 (brs, 1H), 8.31 (s, 1H), 7.40 (d, J=7.2 Hz, 1H), 7.12 (m,3H), 4.42 (t, J=6.8 Hz, 1H), 4.11 (m, 1H), 3.98 (m, 3H), 3.70 (m, 2H),3.61 (m, 1H), 3.41 (m, 1H), 2.96 (m, 1H), 2.80 (m, 1H), 2.21 (m, 1H),1.94 (m, 1H). LCMS (APCI+) m/z 400 [M+H]⁺; Rt=1.52 min.

Example 101

Preparation of4-Amino-2-(4-methylbenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-one,dihydrochloride

Step 1: To a solution containing LiHMDS (1.0M, 11.3 mL, 11 mmol) in 40mL of THF under a nitrogen atmosphere at −78 C was added a solution of2-Oxo-pyrrolidine-1-carboxylic acid tert-butyl ester (2 .gg, 11 mmol) in15 mL of THF dropwise over 5 minutes. After complete addition, thereaction was allowed to stir at −78° C. for 45 minutes, followed by theaddition of a solution containing 4-methyl benzyl bromide (2.1 g, 11mmol) in 15 mL of THF dropwise over 5 minutes. The reaction was allowedto stir at −78 C for 1 hour then warmed to 0 C and stirred for 1 hour.The mixture was quenched with 36 mL of 3M LiOH and allowed to stir atroom temperature overnight. The reaction was diluted with water andwashed with ether. The aqueous phase was acidified with 1N HCl andextracted with DCM. The organic phase was dried over magnesium sulfate.Filtration and removal of solvent gave4-tert-Butoxycarbonylamino-2-(4-methylbenzyl)-butyric acid (1.61 g,49%.) LCMS (APCI—) m/z 306 [M-Boc−H]⁻; Rt: 2.14 min.

Step 2: To a solution containing4-tert-Butoxycarbonylamino-2-(4-methylbenzyl)-butyric acid (0.24 g, 0.77mmol) in 25 mL of DMF under a nitrogen atmosphere was added EDCI (0.16g, 0.84 mmol), HOBT (130 mg, 0.84 mmol) and NMM (0.28 g, 2.8 mmol.)After stirring at room temperature for 15 minutes,4-piperazin-1-yl-quinazoline (200 mg, 0.93 mmol) was added and thereaction allowed to stir at room temperature. The reaction was dilutedwith ethyl acetate and washed with water, saturated sodium bicarbonateand water. The organic phase was dried over magnesium sulfate.Filtration, removal of solvent and purification of the residue viabiotage eluting with 10% MeOH/DCM gave[3-(4-Methyl-benzyl)-4-oxo-4-(4-quinazolin-4-yl-piperazin-1-yl)-butyl]-carbamicacid tert-butyl ester (0.225 g, 64%) as a white solid. LCMS (APCI+) m/z504 [M+H]⁺; Rt: 3.03 min. ¹H NMR (CDCl₃, 400 MHz). 8.69 (1H, s), 7.92(1H, d, J 8.3 Hz), 7.76-7.71 (2H, m), 7.45 (1H, t, J 7.8 Hz), 7.06 (4H,m), 4.53 (1H, br. s), 3.96-3.76 (2H, m), 3.63-3.54 (2H, m), 3.47-3.40(2H, m), 3.24-2.84 (6H, m), 2.76-2.70 (1H, m), 2.27 (3H, s), 2.09-2.00(1H, m), 1.73-1.65 (1H, m), 1.40 (9H, s.)

Step 3: A mixture containing[3-(4-Methyl-benzyl)-4-oxo-4-(4-quinazolin-4-yl-piperazin-1-yl)-butyl]-carbamicacid tert-butyl ester (0.22 g, 0.44 mmol) in 10 mL of DCM and mL of 4NHCin dioxane was allowed to stir at room temperature under a nitrogenatmosphere overnight. The reaction was concentrated under reducedpressure. The residue was dissolved in methanol and ether added toprecipitate the product. The solids were filtered and dried to afford4-Amino-2-(4-methylbenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-butan-1-onedihydrochloride (167 mg.) LCMS (APCI+) m/z 404 [M+H]⁺; Rt: 1.87 min. ¹HNMR (D2, 400 MHz). 8.45 (1H, s), 7.87 (2H, t, J 8.1 Hz), 7.65-7.57 (2H,m), 7.01 (4H, s), 4.18-4.12 (1H, m), 3.98-3.92 (1H, m), 3.82-3.71 (2H,m), 3.52-3.37 (3H, m), 3.25-2.76 (6H, m), 2.64-2.54 (1H, m), 2.02 (3H,s), 1.98-1.80 (1H, m.)

Example 102

Preparation of(2R,3′R)-2-(3′-Amino-pyrrolidin-1-yl)-2-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-ethanonetrihydrochloride and(2S,3′R)-2-(3′-Amino-pyrrolidin-1-yl)-2-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-ethanonetrihydrochloride

Step 1: To a solution of glyoxylic acid monohydrate (1.0 g, 10.9 mmol)and (3R)-3-Boc-amino-pyrrolidine (2.06 g, 11.1 mmol) in 65 mL DCE wasadded 3,4-dichlorophenyl boronic acid (2.11 g, 11.1 mmol). The reactionmixture was heated to reflux and stirred 16 hours, after which it wascooled to room temperature, diluted with DCM, and extracted with 1MNa₂CO₃. The basic aqueous layer was extracted with EtOAc, and thecombined extracts were washed with 1N NaOH, dried (Na₂SO₄), filtered,and concentrated. The resulting residue was dissolved in minimal DCM,and the product was triturated by the addition of ether. The resultingsolids were isolated by vacuum filtration, washed with ether, and driedin vacuo to give one diastereomer of(3′R)-(3′-Boc-amino-pyrrolidin-1-yl)-(3,4-difluorophenyl)-acetic acid(1.53 g, 36%) as a tan powder, which will be referred to asDiastereomer 1. ¹H NMR (DMSO-d6, 400 MHz) δ 7.63 (s, 1H), 7.45 (d, J=8.2Hz, 1H), 7.33 (d, J=8.0 Hz, 1H), 7.09 (d, J=6.4 Hz, 0.2H), 6.95 (d,J=6.4 Hz, 0.8H), 3.92-3.78 (m, 1H), 3.51 (s, 0.25H), 3.49 (s, 0.75H),3.40-3.30 (m, 1H), 2.77-2.69 (m, 0.2H), 2.64-2.53 (m, 1.8H), 2.42-2.33(m, 1H), 2.31-2.17 (m, 1H), 2.01-1.88 (m, 1H), 1.58-1.46 (m, 1H), 1.36(s, 9H). LCMS (APCI)+m/z 389 [M+H]₊; HPLC Rt 2.17 min.

The basic aqueous layer from above was carefully acidified to about pH6.5 with solid KHSO₄ until CO₂ evolution ceased. The oily mixture wasthen extracted with EtOAc, and the combined extracts were dried(Na₂SO₄), filtered, and concentrated. The resulting residue wasdissolved in minimal DCM, and the product was triturated by the additionof ether. The resulting solids were isolated by vacuum filtration,washed with ether, and dried in vacuo to give the other diastereomer of(3′R)-(3′-Boc-amino-pyrrolidin-1-yl)-(3,4-difluorophenyl)-acetic acid(0.67 g, 16%) as a tan powder, which will be referred to as Diastereomer2. ¹H NMR (DMSO-d6, 400 MHz) δ 7.68 (s, 1H), 7.67-7.58 (m, 1.25H),7.46-7.37 (m, 1H), 7.13-7.03 (m, 0.75H), 4.20 (d, J=6.6 Hz, 1H), 3.97(br s, 1H), 3.11-3.00 (m, 0.5H), 2.90-2.76 (m, 1.5H), 2.74-2.63 (m,0.5H), 2.63-2.52 (m, 1H), 2.47-2.37 (m, 0.5H), 2.13-1.97 (m, 1H),1.74-1.60 (m, 1H), 1.36 (s, 9H). LCMS (APCI)+m/z 389 [M+H]₊; HPLC Rt2.17 min.

Step 2, (Diastereomer 1): To a solution of 4-piperazin-1-ylquinazoline(30 mg, 0.14 mmol) and Diastereomer 1 (65 mg, 0.17 mmol) in 1.2 mL 3:1DCM:THF were added successively HOBt.H₂O (21 mg, 0.14 mmol) and DCC (34mg, 0.17 mmol). The reaction mixture was stirred at room temperature 3.5hours, after which it was diluted with DCM, vacuum filtered throughcompressed celite, and rinsed with DCM. The filtrate was then stirredwith 2N NaOH and extracted with DCM. The combined extracts were dried(Na₂SO₄), filtered and concentrated. The crude was purified on silica(1:4 DCM:ethyl acetate to 30:1 DCM:MeOH gradient) to give(3′R)-2-(3′-Boc-amino-pyrrolidin-1-yl)-2-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-ethanone.This was then dissolved in 1.2 mL dioxane, and 1.5 mL 4M HCl/dioxane wasadded. The resulting suspension was stirred at room temperature 16hours, after which it was concentrated to dryness. The solids weredissolved in minimal MeOH, and the product was triturated with ether.The resulting solids were isolated by filtration through a frittedfunnel with nitrogen pressure, rinsed with ether, and dried in vacuo togive one diasteomer of(3′R)-2-(3′-amino-pyrrolidin-1-yl)-2-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-ethanonetrihydrochloride (67 mg, 81%) as a pale yellow powder. ¹H NMR (D₂O, 400MHz) δ 8.45 (s, 1H), 7.91 (d, J=8.0 Hz, 1H), 7.89 (d, J=7.8 Hz, 1H),7.67 (d, J=8.4 Hz, 1H), 7.59 (d, J=7.8 Hz, 1H), 7.56 (s, 1H), 7.51 (d,J=8.4 Hz, 1H), 7.33 (d, J=8.4 Hz, 1H), 5.54 (s, 0.75H), 5.52 (s, 0.25H),4.25-3.95 (m, 5H), 3.77-3.35 (m, 7H), 3.26-3.06 (m, 1H), 2.60-2.40 (m,1H), 2.19-1.94 (m, 1H). LCMS (APCI)+m/z 485 [M+H]⁺; HPLC Rt 1.71 min.

Step 2, Diastereomer 2: 4-piperazin-1-ylquinazoline was acylated withDiastereomer 2 following Step 2 for Diastereomer 1 above, to furnish theother diastereomer of(3′R)-2-(3-amino-pyrrolidin-1-yl)-2-(3,4-dichlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-ethanonetrihydrochloride (60 mg, 72%) as a pale yellow powder. ¹H NMR (D₂O, 400MHz) δ 8.45 (s, 1H), 7.91 (d, J=7.8 Hz, 1H), 7.89 (d, J=7.8 Hz, 1H),7.67 (d, J=8.4 Hz, 1H), 7.59 (d, J=8.0 Hz, 1H), 7.55 (s, 1H), 7.49 (d,J=8.2 Hz, 1H), 7.32 (d, J=7.6 Hz, 1H), 5.46 (d, J=12.7 Hz, 1H),4.24-3.95 (m, 5H), 3.82-3.44 (m, 6H), 3.40-3.28 (m, 0.5H), 3.20-3.10 (m,1H), 3.10-2.98 (m, 0.5H), 2.56-2.36 (m, 1H), 2.15-1.92 (m, 1H). LCMS(APCI)+m/z 485 [M+H]₊; HPLC Rt 1.69 min.

Example 103

The Preparation of(2R)-2-amino-3-phenyl-1-(4-pyrido[2,3-d]pyrimidin-4-yl-piperazin-1-yl)-propan-1-onetrihydrochloride

Step 1: 2-aminonicotinic acid (7 g) and formamide (22.8 g) were heatedat 167 C (internal temperature) for 2.5 hours. After cooling, the solidwas recrystallized from 100 mL of hot water to givePyrido[2,3-d]pyrimidin-4-ol as pale yellow powder (5.2 g, 69.7%). ¹H NMR(DMSO, 400 MHz) δ 12.60 (br, 1H), 8.90 (br, 1H), 8.50 (m, 1H), 8.37 (s,1H), 7.50 (m, 1H). R_(t) 0.87 min. MS (ESI+) [M+H]⁺ 148.

Step 2: The Pyrido[2,3-d]pyrimidin-4-ol (3 g) in POCl₃ (45 mL) wasstirred at reflux for 3 hours. The excess POCl₃ was removed. The residuewas added 10 mL of cold water and extracted with EtOAc (2×30 mL). Thecombined organic layer was washed with brine and dried over sodiumsulfate. After removal of solvent, it gave4-chloropyrido[2,3-d]pyrimidine as yellow solid (0.3 g, 7.6%). ¹H NMR(CDCl₃, 400 MHz) δ 9.38 (br, 1H), 9.31 (s, 1H), 8.62 (d, 1H), 7.75 (m,1H).

Step 3: The 4-chloro-pyrido[2,3-d]pyrimidine (0.3 g) and piperazine (1.6g) in Ethanol (10 mL) was refluxed for 1 hour. The solvent was removedand 50 mL of toluene was added. The toluene was removed in vacuo. Theresulting solid was used directly for the next step withoutpurification. R_(t) 1.93 min. MS (ESI+) [M+H]⁺ 216.

Step 4: DIEA (0.74 mL) and HBTU (1.3 g) was added to the solution of(2R)-2-tert-butoxycarbonylamino-3-phenyl-propionic acid (0.092 g) in THF(5 mL) at 0° C. The mixture was stirred at room temperature for 20minutes, and then 4-piperazin-1-yl-pyrido[2,3-d]pyrimidinene (0.31 g)was added. The reaction was stirred at room temperature for 1 hour. 20mL of EtOAc was added and the organic layer was separated. The aqueouslayer extracted with EtOAc (20 mL). The combined organic layer waswashed with saturated sodium bicarbonate (20 mL) and dried over sodiumsulfate. After removal of solvent, the residue was purified by flashchromatography (10:1=DCM:MeOH) to give[1-benzyl-2-oxo-2-(4-pyrido[2,3-d]pyrimidin-4-yl-piperazin-yl)-ethyl-carbamicacid tert-butyl ester as white foam solid (0.392 g, 58.9%). ¹H NMR(CDCl₃, 400 MHz) δ 9.11 (br, 1H), 8.92 (s, 1H), 8.15 (d, 1H), 7.40 (m,1H), 7.20-7.33 (m, 5H), 5.42 (m, 1H), 4.85 (m, 1H), 3.78-3.82 (m, 2H),3.61-3.70 (m, 2H), 3.50-3.58 (m, 2H), 3.15-3.20 (m, 1H), 3.08-3.12 (m,2H), 1.42 (s, 9H). R_(t) 2.23 min. MS (ESI+) [M+H]⁺ 463.

Step 5:[1-benzyl-2-oxo-2-(4-pyrido[2,3-d]pyrimidin-4-yl-piperazin-yl)-ethyl-carbamicacid tert-butyl ester (0.046 mg) was dissolved in DCM (5 mL) andHCl/dioxane (0.5 mL) was added. The suspension was stirred at roomtemperature for 3 hours, after which it was concentrated to give(2R)-2-amino-3-phenyl-1-(4-pyrido[2,3-d]pyrimidin-4-yl-piperazin-1-yl)-propan-1-onetrihydrochloride (0.031 g, 86%). R_(t) 1.55 min. MS (ESI+) [M+H]⁺ 363.

Example 104

Preparation of5-amino-2-(4-chlorobenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-pentan-1-onedihydrochloride

Step 1: To a −78° C. solution of LHMDS (1.0M, 5.3 mL, 5.3 mmol) in 10 mLTHF was added by syringe a solution of Boc-2-piperidone (1.0 g, 5.0mmol) in 8 mL THF. The reaction mixture was stirred at −78° C. for 1hour, after which a 0° C. solution of 4-chlorobenzyl bromide (1.1 g, 5.3mmol) in 5 mL THF was added quickly by syringe. The reaction mixture wasstirred 1 hour at −78° C., warmed to 0° C., stirred another 1 hour, thenquenched with 17 mL 3M LiOH solution. The reaction mixture was thenstirred 15 hours at room temperature, after which it was diluted withH₂O and washed with ether. The aqueous layer was acidified with solidKHSO₄, extracted with DCM, and the extracts were dried (Na₂SO₄),filtered, and concentrated to give5-Boc-amino-2-(4-chlorobenzyl)-pentanoic acid (1.0 g, 60%) as a clear,colorless syrup. LCMS (APCI—) m/z 340 [M−H]⁻; Rt: 2.37 min.

Step 2: A solution of EDCI (100 mg, 0.54 mmol), HOBt.H₂O (82 mg, 0.54mmol), 5-Boc-amino-2-(4-chlorobenzyl)-pentanoic acid (170 mg, 0.50mmol), and TEA (190 μL, 1.4 mmol) in 3 mL DMF was stirred 10 minutes,and solid 4-piperazin-1-yl-quinazoline dihydrochloride (130 mg, 0.45mmol) was added. The reaction mixture was stirred at room temperature 15hours, after which water was added. The reaction mixture was extractedwith DCM, and combined extracts washed with sat NaHCO₃, dried (Na₂SO₄),filtered, and concentrated. The crude was purified on silica gel (1:1 to1:9 DCM:EtOAc) to give5-Boc-amino-2-(4-chlorobenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-pentan-1-one.LCMS (APCI+) m/z 538 [M+H]⁺; Rt: 3.12 min.

Step 3: To a solution of5-Boc-amino-2-(4-chlorobenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-pentan-1-onein 1.5 mL dioxane was added 2.5 mL 4M HCl/dioxane. The resultingsuspension was stirred at room temperature 17 hours, after which it wasconcentrated to dryness. The solids were dissolved in minimal MeOH, andthe product was triturated by the addition of ether. The solids wereisolated by filtration through a fritted funnel with nitrogen pressure,rinsed with ether, and dried in vacuo to give5-amino-2-(4-chlorobenzyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-pentan-1-onedihydrochloride (110 mg, 46%) as a white powder. ¹H NMR (CD₃OD, 400 MHz)δ 8.71 (1H, d, J 2.0 Hz), 8.20 (1H, d, J 8.7 Hz), 8.05 (1H, t, J 7.8Hz), 7.84-7.75 (2H, m), 7.31-7.22 (4H, m), 4.40-4.30 (1H, m), 4.29-4.20(1H, m), 4.12-4.02 (1H, m), 3.92-3.66 (4H, m), 3.53-3.42 (1H, m),3.29-3.21 (1H, m), 2.97-2.81 (4H, m), 1.87-1.62 (4H, m). LCMS (APCI+)m/z 438 [M+H]⁺; Rt: 1.98 min.

Example 105

Preparation of2-(2-Amino-ethoxy)-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-onedihydrochloride

The PS-CDI (175 mg, 0.181 mmol), HOBt monohydrate (214 mg, 0.140 mmol),4-piperazin-1-yl-quinazoline dihydrochloride (52 mg, 0.181 mmol), and2-(2-tert-butoxycarbonylamino-ethoxy)-3-(4-chlorophenyl)-propionic acid(48 mg, 0.140 mmol; prepared by alkylation of(2-Boc-amino-ethoxy)-acetic acid ethyl ester with 4-chlorobenzylbromideaccording to procedures described in the literature: Nizal S.Chandrakumar et al. J. Med. Chem. 1992, 35, 2928-2938) weresuspended/dissolved in 2.5 mL of chloroform (plus 3-5 drops THF). Themixture was shaken at room temperature overnight, then treated withMP-CO₃ (330 mg, 0.838 mmol) for two hours. The mixture was vacuumfiltered, rinsed with chloroform, and concentrated in vacuo. The residuewas purified on silica gel (1:19 DCM:EtOAc) to afford2-(2-Boc-amino-ethoxy)-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-one.This material was dissolved in 1.0 mL of 1,4-dioxane and treated with1.5 mL of 4M HCl in 1,4-dioxane affording slow precipitation. Themixture was sonicated briefly and stirred at room temperature overnightto completion. The resulting suspension was concentrated in vacuo, andthe solids were suspended in diethyl ether, filtered under nitrogenpressure, and dried in vacuo to afford2-(2-amino-ethoxy)-3-(4-chlorophenyl)-1-(4-quinazolin-4-yl-piperazin-1-yl)-propan-1-onedihydrochloride as a yellow powder (13 mg, 18%). LCMS (APCI+) m/z 440[M+H]⁺. HPLC R_(t)=1.75 min.

Example 106

Preparation of(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(3-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1: To a stirred solution of LDA (9.85 mL, 1.5 M, 15.8 mmol) in THF(20 mL) was added dropwise a solution of 4,6-dichloropyrimidine (2.00 g,13.4 mmol) in THF (12 mL) at −78° C. After stirring for 1.5 hours, asolution of cyclopropanecarbaldehyde (1.05 g, 15.0 mmol) in THF (10 mL)was added dropwise. The solution was stirred at −78° C. for 1 hour andthen quenched by addition of water (10 mL). The reaction mixture wasallowed to warm to room temperature and partitioned between EtOAc andwater. The organic layer was washed with brine, dried and concentrated.The residue was purified by column chromatography (hexanes/EtOAc, 3:1)to give Cyclopropyl-(4,6-dichloro-pyrimidin-5-yl)-methanol (2.36 g, 80%)as a yellow oil. ¹H NMR (CDCl₃, 400 MHz) δ 8.70 (s, 1H), 4.56 (m, 1H),2.63 (d, J=8.0 Hz, 1H), 1.70 (m, 1H), 0.76 (m, 1H), 0.55 (m, 2H).

Step 2: To a vigorously stirred solution ofCyclopropyl-(4,6-dichloro-pyrimidin-5-yl)-methanol (0.84 g, 3.8 mmol) inanhydrous acetone (12 mL) was added portionwise chromium (VI) oxide (1.2g, 12 mmol) at 0° C. The mixture was stirred at 0° C. for 30 minutes.The excess of the oxidizing agent was destroyed by the addition ofisopropanol (2 mL). After stirring for 15 minutes, the reaction mixturewas poured into saturated NaHCO₃ solution and filtered through Celite.The filtrate was extracted with EtOAc. The combined organic layers werewashed with brine, dried and concentrated. The residue was purified bycolumn chromatography (hexanes:EtOAc, 6:1) to giveCyclopropyl-(4,6-dichloro-pyrimidin-5-yl)-methanone (0.80 g, 96%) as acolorless oil. ¹H NMR (CDCl₃, 400 MHz) δ 8.83 (s, 1H), 2.26 (m, 1H),1.46 (m, 2H), 1.26 (m, 2H).

Step 3: A mixture of Cyclopropyl-(4,6-dichloro-pyrimidin-5-yl)-methanone(0.75 g, 3.5 mmol), anhydrous hydrazine (0.13 mL, 4.1 mmol) and THF (35mL) was stirred at room temperature for 4 hours. The reaction waspartitioned between water and EtOAc. The organic layer was washed withbrine, dried and passed through a short silica gel pad to give4-Chloro-3-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine (0.50 g, 74%) as awhite solid. ¹H NMR (CDCl₃, 400 MHz) δ 11.87 (s, 1H), 8.80 (s, 1H), 2.55(m, 1H), 1.15 (m, 2H), 1.14 (m, 2H).

Step 4:4-(3-Cyclopropyl-11H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester was prepared by the procedures described inExample 40, Step 1, substituting 4-chloro-5-iodopyrimidine with4-Chloro-3-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidine. LCMS (APCI+) m/z345 [M+H]⁺; Rt=2.52 min.

Step 5: 3-Cyclopropyl-4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidinedihydrochloride was prepared by the procedures described in Example 34,Step 3, substituting(2R)-{1-(4-Chlorobenzyl)-2-[4-(1H-indazol-5-yl)-piperazin-1-yl]-2-oxo-ethyl}-carbamicacid tert-butyl ester with4-(3-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazine-1-carboxylicacid tert-butyl ester. ¹H NMR (DMSO-d₆, 400 MHz) δ 8.58 (s, 1H), 4.60(m, 4H), 3.53 (m, 4H), 2.36 (m, 1H), 1.31 (m, 2H), 1.14 (m, 2H). LCMS(APCI+) m/z 245 [M+H]⁺; Rt=1.02 min.

Step 6:(2R)-2-Amino-3-(4-chlorophenyl)-1-[4-(3-cyclopropyl-1H-pyrazolo[3,4-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride was prepared by substituting4-piperazin-1-yl-6,7,8,9-tetrahydro-5H-1,3,9-triaza-fluorenedihydrochloride with3-cyclopropyl-4-piperazin-1-yl-1H-pyrazolo[3,4-d]pyrimidinedihydrochloride in Example 81, Step 3. ¹H NMR (CD₃OD, 400 MHz) δ 8.48(s, 1H), 7.40 (d, J=8.4 Hz, 2H), 7.34 (d, J=8.4 Hz, 2H), 4.77 (m, 1H),4.24 (m, 3H), 3.87 (m, 2H), 3.74 (m, 2H), 3.25 (m, 1H), 3.16 (m, 2H),2.27 (m, 1H), 1.28 (m, 2H), 1.10 (m, 2H). LCMS (APCI+) m/z 426, 428[M+H]⁺; Rt=1.88 min.

Example 107

Preparation of2-(3,4-Dichlorophenyl)-3-(1H-imidazol-4-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onedihydrochloride

Step 1:4-[2-(3,4-Dichlorophenyl)-2-methoxycarbonyl-ethyl]-imidazole-1-carboxylicacid tert-butyl ester was prepared by the procedures described inExample 78, Step 1, substituting 3-tert-butoxycarbonylamino-propionicacid tert-butyl ester with (3,4-Dichlorophenyl)-acetic acid methyl esterand substituting 4-Bromo-1-bromomethyl-2-fluoro-benzene with4-Bromomethyl-imidazole-1-carboxylic acid tert-butyl ester (preparedfrom 4(5)-hydroxymethylimidazole hydrochloride according to theliterature: J. Med. Chem. 1997, 40, 2208). ¹H NMR (CDCl₃, 400 MHz) δ7.96 (s, 1H), 7.42 (s, 1H), 7.38 (d, J=8.0 Hz, 1H), 7.16 (d, J=8.0 Hz,1H), 7.02 (s, 1H), 4.08 (t, J=7.6 Hz, 1H), 3.66 (s, 3H), 3.32 (dd,J=14.4 Hz, J=8.4 Hz, 1H), 2.92 (dd, J=14.4 Hz, J=6.8 Hz, 1H). LCMS(APCI+) m/z 299, 301, 303 [M-Boc+H]⁺; Rt=3.72 min.

Step 2: 2-(3,4-Dichlorophenyl)-3-(1H-imidazol-4-yl)-propionic acid wasprepared by the procedures described in Example 78, Step 2, substituting3-tert-Butoxycarbonylamino-2-(4-trifluoromethylbenzyl)-propionic acidethyl ester with4-[2-(3,4-Dichlorophenyl)-2-methoxycarbonyl-ethyl]-imidazole-1-carboxylicacid tert-butyl ester. ¹H NMR (CD₃OD, 400 MHz) δ 8.25 (s, 1H), 7.51 (s,1H), 7.42 (d, J=8.0 Hz, 1H), 7.26 (d, J=8.0 Hz, 1H), 7.00 (s, 1H), 3.84(m, 1H), 3.38 (m, 1H), 3.00 (dd, J=14.4 Hz, J=6.8 Hz, 1H). LCMS (APCI+)m/z 285, 287, 289 [M+H]⁺; Rt=1.54 min.

Step 3:2-(3,4-Dichlorophenyl)-3-(1H-imidazol-4-yl)-1-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-piperazin-1-yl]-propan-1-onewas prepared by substituting 5-piperazin-1-yl-1H-indazole with4-piperazin-1-yl-7H-pyrrolo[2,3-d]pyrimidine dihydrochloride andsubstituting (D)-Boc-4-chlorophenylalanine with2-(3,4-Dichlorophenyl)-3-(1H-imidazol-4-yl)-propionic acid in ExampleB-1, Step 2. The free amine was converted to HCl salt by treatment withHCl in Ether. ¹H NMR (CD₃OD, 400 MHz) δ 8.76 (s, 1H), 8.32 (s, 1H), 7.52(m, 2H), 7.38 (s, 1H), 7.29 (m, 2H), 6.92 (s, 1H), 4.58 (m, 1H),3.70-4.20 (m, 7H), 3.63 (m, 1H), 3.43 (m, 1H), 3.16 (m, 1H). LCMS(APCI+) m/z 470, 472, 474 [M+H]⁺; Rt=2.15 min.

The words “comprise,” “comprising,” “include,” “including,” and“includes” when used in this specification and in the following claimsare intended to specify the presence of stated features, integers,components, or steps, but they do not preclude the presence or additionof one or more other features, integers, components, steps, or groups.

What is claimed is:
 1. A compound including resolved enantiomers,diastereomers, solvates and pharmaceutically acceptable salts thereof,said compound comprising Formula I:A-L-CR  (I) where: CR is heteroaryl, wherein said heteroaryl isoptionally substituted with one or more groups selected from halogen,hydroxyl, cyano, nitro, azido, —NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴,—S(O)NR²¹R²², —C(O)R²¹, —C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, —NR²¹C(O)OR²⁴,—NR²¹C(═NR²¹)NR²²R²³, —NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴,—SO₂R²⁴, —NR²¹R²², —NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³, —OR²¹, C₁-C₄alkyl, C₁-C₆ heteroalkyl, C₂-C₆ alkenyl, C₂-C₆ heteroalkenyl, C₂-C₆alkynyl, C₂-C₆ heteroalkynyl, C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl,aryl, heteroaryl, arylalkyl and heteroarylalkyl, wherein any of saidalkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl,cycloalkyl, heterocycloalkyl, aryl, heteroaryl, arylalkyl andheteroarylalkyl are further optionally substituted with one or moregroups selected from halogen, hydroxyl, cyano, nitro, azido,fluoromethyl, difluoromethyl, trifluoromethyl, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₁-C₆ heteroalkyl, C₂-C₆ heteroalkenyl, C₂-C₆heteroalkynyl, C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl, —SR²¹,—S(O)R²⁴, —SO₂R²⁴, —C(O)R²¹, C(O)OR²¹, —C(O)NR²¹R²², —NR²¹R²² and —OR²¹;L is selected from:

where R¹² is hydrogen, halogen, hydroxy, cyano, nitro, amino, azido,C₁-C₅ alkyl, C₂-C₅ alkenyl, C₂-C₅ alkynyl, C₁-C₅ heteroalkyl, C₂-C₅heteroalkenyl or C₂-C₅ heteroalkynyl, wherein any of said alkyl,alkenyl, alkynyl, heteroalkyl, heteroalkenyl, and heteroalkynyl areoptionally substituted with one or more groups independently selectedfrom halogen, cyano, nitro, amino, azido, C₁-C₄ alkyl, fluoromethyl,difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy, difluoromethoxyand trifluoromethoxy;

A is W is N or CR¹⁵, provided that when L is a substituted orunsubstituted piperazinylene, W must be CR¹⁵; G is hydrogen, alkyl,heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,heterocycloalkyl, aryl or heteroaryl, wherein any of said alkyl,heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,heterocycloalkyl, aryl or heteroaryl is optionally substituted with oneor more groups selected from halogen, hydroxyl, cyano, amino, nitro,azido, —NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹,—C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, —NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³,—NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴, —SO₂R²⁴, —NR²¹R²²,—NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³, —OR²¹, C₁-C₄ alkyl, C₁-C₄heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄heteroalkynyl, cycloalkyl, heterocycloalkyl aryl and heteroaryl; B¹ andB² are independently absent or C₁-C₄ alkylene, C₁-C₄ heteroalkylene,C₂-C₄ alkenylene, C₂-C₄ heteroalkenylene, C₂-C₄ alkynylene, C₂-C₄heteroalkynylene, C₃-C₆ cycloalkylene, and C₃-C₆ heterocycloalkylene,wherein any of said alkylene, heteroalkylene, alkenylene,heteroalkenylene, alkynylene, heteroalkynylene, cycloalkylene orheterocycloalkylene is optionally substituted with one or more groupsindependently selected from halogen, hydroxyl, cyano, nitro, azido,C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl,C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, NR²¹R²² and OR²¹; R²¹, R²² and R²³independently are hydrogen, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₁-C₆ heteroalkyl, C₂-C₆ heteroalkenyl, C₂-C₆ heteroalkynyl, C₃-C₆cycloalkyl, C₃-C₆ heterocycloalkyl, aryl, arylalkyl, heteroaryl orheteroarylalkyl; R²⁴ is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆heteroalkyl, C₂-C₆ heteroalkenyl, C₂-C₆ heteroalkynyl, C₃-C₆ cycloalkyl,C₃-C₆ heterocycloalkyl, aryl, arylalkyl, heteroaryl or heteroarylalkyl;or any two of R²¹, R²², R²³ or R²⁴ together with the atom(s) to whichthey are attached form a 4 to 10 membered carbocyclic, aryl, heteroarylor heterocyclic ring, wherein any of said carbocyclic, aryl, heteroarylor heterocyclic rings are optionally substituted with one or more groupsindependently selected from halogen, cyano, nitro, trifluoromethyl,difluoromethoxy, trifluoromethoxy, azido, aryl, heteroaryl, arylalkyl,heteroarylalkyl, heterocyclyl, and heterocyclylalkyl; R¹³ and R¹⁴ areindependently hydrogen, hydroxyl, cyano, C₁-C₆ alkyl, C₁-C₆ heteroalkyl,C₂-C₆ alkenyl, C₂-C₆ heteroalkenyl, C₂-C₆ alkynyl, C₂-C₆ heteroalkynyl,C₁-C₆ cycloalkyl, C₁-C₆ heterocycloalkyl, aryl, heteroaryl, arylalkyl,heteroarylalkyl, —C(O)R²¹, C(O)OR²¹, C(═NR²¹)NR²²R²³ or —SO₂R²⁴, whereinany of said alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, heteroalkyl, heteroalkenyl, heteroalkynyl, arylalkyl orheteroarylalkyl is optionally substituted with one or more groupsindependently selected from halogen, hydroxyl, cyano, amino, nitro,azido, —NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹,—C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, —NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³,—NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴, —SO₂R²⁴, —NR²¹R²²,—NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³, —OR²¹, C₁-C₄ alkyl, C₁-C₄heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄heteroalkynyl, cycloalkyl, heterocycloalkyl aryl and heteroaryl; or R¹³and R¹⁴ together with the atoms to which they are attached form a 4 to10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring, whereinany of said carbocyclic, aryl, heteroaryl and heterocyclic rings areoptionally substituted with one or more groups independently selectedfrom halogen, cyano, nitro, azido, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl,NR²¹R²² and OR²¹; or R¹³ and an atom of B² together with N form a 4 to10 membered carbocyclic, aryl, heteroaryl or heterocyclic ring, whereinany of said carbocyclic, aryl, heteroaryl and heterocyclic rings areoptionally substituted with one or more groups independently selectedfrom halogen, cyano, nitro, azido, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl,NR²¹R²² and OR²¹; R¹⁵ is hydrogen, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl or C₂-C₄ heteroalkynyl,wherein any of said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynylor heteroalkynyl is optionally substituted with one or more groupsindependently selected from halogen, hydroxyl, cyano, nitro, azido,NR²¹R²² and OR²¹; or R¹³ and R¹⁵ together with atoms to which they areattached form a 3 to 10 membered carbocyclic, aryl, heteroaryl orheterocyclic ring, wherein any of said carbocyclic, aryl, heteroaryl andheterocyclic rings are optionally substituted with one or more groupsindependently selected from halogen, cyano, nitro, azido, C₁-C₄ alkyl,C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl,C₂-C₄ heteroalkynyl, NR²¹R²² and OR²¹; or, when W is CR¹⁵, R¹⁵ and anatom of B¹ or B² together with C, form a 3 to 10 membered carbocyclic,aryl, heteroaryl or heterocyclic ring, wherein any of said carbocyclic,aryl, heteroaryl and heterocyclic rings are optionally substituted withone or more groups independently selected from halogen, cyano, nitro,azido, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, NR²¹R²² and OR²¹. 2.The compound of claim 1, where CR is selected from:

where X is N or CR¹; Y is CR² or N, provided that when X is N, Y must beCR²; Z is CR³R^(3a) or NR^(2a), provided that when X is N, Z must beCR³; D¹, D², D³ and D⁴ are independently CR⁴ or N, provided that no morethan two of D¹, D², D³ or D⁴ are N; - - - - - is an optional doublebond; D⁵ is CR⁵R^(5a), NR^(2a), O or S, provided that when D⁵ is O or S,D⁸ must be C, D⁷ must be CR⁷ or N, and either (i) Q must be CR⁶ orCR⁶R^(6a) or (ii) D⁷ must be CR⁷ or CR⁷R^(7a); Q is CR⁶, N or C═O,provided that either (w) when Q is N, one of D⁵, D⁷ and D⁸ must be C, or(x) when Q is C═O, D⁵ must be CR⁵ or N, D⁷ must be CR⁷ or N, and D⁸ mustbe C; D⁷ is CR⁷, N, O or S, provided that when D⁷ is O or S, D⁸ must beC, D⁵ must be CR⁵ or N, and either (y) Q must be CR⁶, or (z) D⁵ must beCR⁵; D⁸ is C or N, provided that when D⁸ is N, D⁵ must be CR⁵R^(5a) andQ must be CR⁶ or CR⁶R^(6a); either K or M is carbonyl, provided thatboth K and M are not carbonyl; R¹, R⁵, R^(5a) and R⁸ are independentlyhydrogen, halogen, hydroxyl, cyano, nitro, azido, fluoromethyl,difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy,difluoromethoxy, trifluoromethoxy, amino, aminomethyl, dimethylamino,aminoethyl, diethylamino or ethoxy; R² is hydrogen, halogen, hydroxyl,cyano, nitro, amino, azido, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, C₁-C₆cycloalkyl, C₁-C₆ heterocycloalkyl, C₁-C₆ aryl, or C₁-C₆ heteroaryl,wherein any of said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl arefurther optionally substituted with one or more groups independentlyselected from halogen, hydroxyl, cyano, nitro, azido, fluoromethyl,difluoromethyl, trifluoromethyl, methoxy, fluoromethoxy,difluoromethoxy, trifluoromethoxy, amino, aminomethyl, dimethylamino,aminoethyl, diethylamino and ethoxy; R^(2a) is hydrogen, hydroxyl,cyano, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl, C₂-C₆ heteroalkynyl, C₃-C₆ cycloalkyl,C₃-C₆ heterocycloalkyl, aryl or heteroaryl, wherein any of said alkyl,heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,heterocycloalkyl, aryl or heteroaryl are optionally substituted with oneor more groups independently selected from halogen, hydroxyl, cyano,nitro, azido fluoromethyl, difluoromethyl, trifluoromethyl, methoxy,fluoromethoxy, difluoromethoxy, trifluoromethoxy, amino, aminomethyl,dimethylamino, aminoethyl, diethylamino or ethoxy; R³ and R^(3a) areindependently hydrogen, halogen, hydroxyl, cyano, nitro, amino azido,C₁-C₆ alkyl, C₁-C₆ heteroalkyl, C₂-C₆ alkenyl, C₂-C₆ heteroalkenyl,C₂-C₆ alkynyl, C₂-C₆ heteroalkynyl, C₁-C₆ cycloalkyl, C₁-C₆heterocycloalkyl, aryl or heteroaryl, wherein any of said alkyl,heteroalkyl, alkenyl, heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl,heterocycloalkyl, aryl or heteroaryl are further optionally substitutedwith one or more groups independently selected from halogen, hydroxyl,cyano, nitro, azido, OR¹, NR¹R², and (C═O)R²; R⁴, R⁶, R^(6a), R⁷, R^(7a)and R¹⁰ are independently hydrogen, hydroxyl, cyano, amino, nitro,azido, alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl,heteroaryl, heteroarylalkyl, —NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴,—S(O)NR²¹R²², —C(O)R²¹, —C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, —NR²¹C(O)OR²⁴,—NR²¹C(═NR²¹)NR²²R²³, —NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴,—SO₂R²⁴, —NR²¹R²², —NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³ or —OR²¹,wherein any of said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl, heteroarylor heteroarylalkyl, is optionally substituted with one or more groupsindependently selected from halogen, hydroxyl, cyano, amino, nitro,azido, —NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹,—C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, —NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³,—NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴, —SO₂R²⁴, —NR²¹R²²,—NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³, —OR²¹, C₁-C₄ alkyl, C₁-C₄heteroalkyl, C₂-C₄ alkenyl, C₂-C₄ heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, andwherein said alkyl, heteroalkyl, alkenyl, heteroalkenyl, alkynyl,heteroalkynyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl mayoptionally be further optionally substituted with one or more groupsindependently selected from halogen, hydroxyl, cyano, amino, nitro,azido, C₁-C₄ alkyl, C₁-C₄ heteroalkyl, C₂-C₄ alkenyl, C₂-C₄heteroalkenyl, C₂-C₄ alkynyl, C₂-C₄ heteroalkynyl, cycloalkyl,heterocycloalkyl, aryl and heteroaryl, —NR²¹R²², and —OR²¹; or R⁶ and R⁷together with the atoms to which they are attached form a 4 to 10membered carbocyclic, aryl, heteroaryl or heterocyclic ring, wherein anyof said carbocyclic, aryl, heteroaryl and heterocyclic rings areoptionally substituted with one or more groups independently selectedfrom halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,trifluoromethoxy, azido, OR¹, NR¹R², cycloalkyl, heterocycloalkyl, aryl,heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl, andheterocyclylalkyl; and R⁹ is hydrogen, halogen, hydroxyl, cyano, nitro,azido, C₁-C₆ alkyl, C₁-C₆ heteroalkyl, C₂-C₆ alkenyl, C₂-C₆heteroalkenyl, C₂-C₆ alkynyl, C₂-C₆ heteroalkynyl, C₁-C₆ cycloalkyl,C₁-C₆ heterocycloalkyl, aryl, heteroaryl, —NR²¹R²², —OR²¹, —NR²¹SO₂R²⁴and —NR²¹C(O)R²², wherein any of said alkyl, heteroalkyl, alkenyl,heteroalkenyl, alkynyl, heteroalkynyl, cycloalkyl, heterocycloalkyl,aryl or heteroaryl are further optionally substituted with one or moregroups independently selected from halogen, hydroxyl, cyano, nitro,azido, OR¹, NR¹R², and (C═O)R².
 3. The compound of claim 2, where CR isselected from:


4. The compound of claim 3, where D⁵ is NH.
 5. The compound of claim 2,where CR is selected from:


6. The compound of claim 2, where L is selected from:


7. The compound of claim 6, where L is:


8. The compound of claim 2, where A is:


9. The compound of claim 2, where A is:

where G is hydrogen, alkyl, cycloalkyl, heterocycloaryl, aryl orheteroaryl, wherein any of said alkyl, cycloalkyl, heterocycloalkyl,aryl or heteroaryl is optionally substituted with one or more groupsselected from halogen, hydroxyl, cyano, amino, nitro, azido,—NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹,—C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, —NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³,—NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴, —SO₂R²⁴, —NR²¹R²²,—NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³, —OR²¹, C₁-C₄ alkyl, C₂-C₄ alkenyl,C₂-C₄ alkynyl, cycloalkyl, heterocycloalkyl aryl and heteroaryl.
 10. Thecompound of claim 6, where A is:

where G is hydrogen, alkyl, cycloalkyl, heterocycloaryl, aryl orheteroaryl, wherein any of said alkyl, cycloalkyl, heterocycloalkyl,aryl or heteroaryl is optionally substituted with one or more groupsselected from halogen, hydroxyl, cyano, amino, nitro, azido,—NR²¹SO₂R²⁴, —SO₂NR²¹R²², —NR²¹S(O)R⁴, —S(O)NR²¹R²², —C(O)R²¹,—C(O)OR²¹, —OC(O)R²¹, —OC(O)OR²¹, —NR²¹C(O)OR²⁴, —NR²¹C(═NR²¹)NR²²R²³,—NR²¹C(O)R²², —C(O)NR²¹R²², —SR²¹, —S(O)R²⁴, —SO₂R²⁴, —NR²¹R²²,—NR²¹C(O)NR²²R²³, —NR²¹C(NCN)NR²²R²³, —OR²¹, C₁-C₄ alkyl, C₂-C₄ alkenyl,C₂-C₄ alkynyl, cycloalkyl, heterocycloalkyl aryl and heteroaryl.
 11. Thecompound of claim 6, wherein A comprises:

where B¹ and B² are, independently, absent or C₁-C₄ alkylene;R^(21a)—R^(21c) are independently H, halogen, CH₃, CF₃, CH₃O, CN, NO₂,NH₂, Ph, OH, or OCH₂Ph; R^(22a), R^(22b), and R²⁴ are independently H,CH₃, or halogen; R^(23a) is H; and R^(23b) is H, CH₃, CH₂NH₂, CH₂NHCH₂,CH₂CH₂NH₂, CH₂CH₂NHCH₂, CH₂CH₂N(CH₂)₂, —(C═O)CH₂NH₂ or —(C═O)CH₂CH₂NH₂;or R^(23a) and R^(23b) are joined to complete a 5 or 6 memberedheterocyclic ring.
 12. The compound of claim 7, wherein A comprises:

where B¹ and B² are, independently, absent or C₁-C₄ alkylene;R^(21a)—R^(21c) are independently H, halogen, CH₃, CF₃, CH₃O, CN, NO₂,NH₂, Ph, OH, or OCH₂Ph; R^(22a), R^(22b), and R²⁴ are independently H,CH₃, or halogen; R^(23a) is H; and R^(23b) is H, CH₃, CH₂NH₂, CH₂NHCH₂,CH₂CH₂NH₂, CH₂CH₂NHCH₂, CH₂CH₂N(CH₂)₂, —(C═O)CH₂NH₂ or —(C═O)CH₂CH₂NH₂;or R^(23a) and R^(23b) are joined to complete a 5 or 6 memberedheterocyclic ring.
 13. The compound of claim 6, wherein A comprises:

wherein R²⁵ and R²⁶ are independently H or CH₃; and R²⁷ is 1-naphthyl,2-naphthyl, 3′-benzylthienyl, 2′-thienyl, 2′-pyridyl, 3′-pyridyl,4′-pyridyl, 4′-thiazolyl, or 3,3-diphenyl.
 14. The compound of claim 7,wherein A comprises:

wherein R²⁵ and R²⁶ are independently H or CH₃; and R²⁷ is 1-naphthyl,2-naphthyl, 3′-benzylthienyl, 2′-thienyl, 2′-pyridyl, 3′-pyridyl,4′-pyridyl, 4′-thiazolyl, or 3,3-diphenyl.
 15. The compound of claim 6,where A is:


16. The compound of claim 7, where A is:


17. The compound of claim 7, where A is a D- or L-amino acid selectedfrom the naturally occurring amino acids, 4-hydroxyproline,hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvaline,beta-alanine, gamma-aminobutyric acid, cirtulline, homocysteine,homoserine, ornithine and methionine sulfone.
 18. The compound of claim17, wherein the amino acid is alanine, phenylalanine, histidine, ortryptophan.
 19. The compound of claim 1, wherein the compound comprises:


20. The compound of claim 1, wherein the compound comprises:

wherein R²⁸ is s H, halogen, CH₃, CF₃, CH₃O, CN, NO₂, NH₂, Ph, OH, orOCH₂Ph.
 21. The compound of claim 1, wherein the compound comprises:

wherein D¹⁶ is O or N; and R²⁹ is H, halogen, CH₃, CF₃, CH₃O, CN, NO₂,NH₂, Ph, OH, or OCH₂Ph.
 22. A method of treating hyperproliferativediseases in a mammal comprising administering a therapeuticallyeffective amount of the compound defined in claim 1 to said mammal. 23.A method of treating hyperproliferative diseases in a mammal comprisingadministering a therapeutically effective amount of the compound definedin claim 2 to said mammal.
 24. A method of treating hyperproliferativediseases in a mammal comprising administering a therapeuticallyeffective amount of the compound defined in claim 3 to said mammal. 25.A method of treating hyperproliferative diseases in a mammal comprisingadministering a therapeutically effective amount of the compound definedin claim 4 to said mammal.
 26. A method of treating hyperproliferativediseases in a mammal comprising administering a therapeuticallyeffective amount of the compound defined in claim 5 to said mammal. 27.A method of treating hyperproliferative diseases in a mammal comprisingadministering a therapeutically effective amount of the compound definedin claim 6 to said mammal.
 28. A method of treating hyperproliferativediseases in a mammal comprising administering a therapeuticallyeffective amount of the compound defined in claim 7 to said mammal. 29.A method of treating hyperproliferative diseases in a mammal comprisingadministering a therapeutically effective amount of the compound definedin claim 8 to said mammal.
 30. A method of treating hyperproliferativediseases in a mammal comprising administering a therapeuticallyeffective amount of the compound defined in claim 9 to said mammal. 31.A method of treating hyperproliferative diseases in a mammal comprisingadministering a therapeutically effective amount of the compound definedin claim 10 to said mammal.
 32. A method of treating hyperproliferativediseases in a mammal comprising administering a therapeuticallyeffective amount of the compound defined in claim 11 to said mammal. 33.A method of treating hyperproliferative diseases in a mammal comprisingadministering a therapeutically effective amount of the compound definedin claim 12 to said mammal.
 34. A composition comprising a compound ofclaim 1 and a pharmaceutically acceptable carrier.
 35. A compositioncomprising a compound of claim 2 and a pharmaceutically acceptablecarrier.
 36. A composition comprising a compound of claim 3 and apharmaceutically acceptable carrier.
 37. A composition comprising acompound of claim 4 and a pharmaceutically acceptable carrier.
 38. Acomposition comprising a compound of claim 5 and a pharmaceuticallyacceptable carrier.
 39. A composition comprising a compound of claim 7and a pharmaceutically acceptable carrier.
 40. A composition comprisinga compound of claim 9 and a pharmaceutically acceptable carrier.
 41. Acompound according to any one of claims 1-21 for use as a medicament.42. A compound according to any one of claims 1-21 for use as amedicament for the treatment of a hyperproliferative disorder.
 43. Theuse of a compound according to any one of claims 1-21 in the manufactureof a medicament for the treatment of a hyperproliferative disorder or aninflammatory condition.